TW201213805A - Lysis buffers for extracting nucleic acids - Google Patents

Abstract

The present teachings relate to the extraction of nucleic acid from solid materials. Provided are useful compositions, methods and kits for obtaining nucleic acids from a solid biological sample or an adhesive material having a biological material adherent or embedded within the adhesive substrate. The extracted nucleic acid can be used in downstream applications such as genotyping, detection, quantification, and identification of the source of the biological material.

Description

201213805 VI. Description of the Invention: [Technical Fields of the Invention] [0001] This application claims the application of the U.S. Provisional Patent Application No. 61,243,136, filed on Sep. 16, 2009, on June 30, 2010. The priority of U.S. Provisional Patent Application No. 61,360,386, and U.S. Provisional Patent Application No. 61,379,346, filed on Sep. 1, 2010. Application Serial No. 10/306, 347, 11/789, 352, and No. 60/334, 029, the entire disclosure of which is incorporated herein by reference.

[0002] 〇 099131421 In general, the teachings relate to the extraction of nucleic acids from solid materials such as bone, teeth and lexical tissue or from biological samples embedded or attached to an adhesive material or denim material. [Prior Art] Extraction of nucleic acids from solid biological materials such as bones and teeth or biological samples containing nucleic acids that are afflicted and/or attached to adhesive and gel-containing materials, and biological materials that are dried or embedded in denim materials There are challenges in processing samples and possible delays in processing samples in forensic laboratories. Forensic samples, missing persons, ancient and degraded samples are also more complex with PCR inhibitors that may be extracted with the extracted nucleic acids. The present teachings provide useful compositions and methods' to obtain nucleic acids (e.g., genomic DNA and rna) from solid biological samples, adhesives having biomaterial attachments or embedded in adherent or denim materials or soil. The extracted nucleic acid can be used for downstream applications such as (genomic typing, detection, quantification, and identification of sources of biological material) using molecular biological steps such as pcR. The lysing solution provided can be used to prepare high amounts of nucleic acids (eg, DNA), as well as biological tissue forms that are extracted from the masculinized tissue or in the knit and/or adhesive adherents and materials as well as denim substrates and materials. No. A0101 Page 3 of 54 ^ 1003014903-0 201213805 ^ADNA. This solution provides a highly efficient method for extracting argon and methods for removing PCR inhibitors, as well as methods for excluding pCR inhibitor extraction. In addition, the procedure for extracting and purifying nucleic acids is fully automated (using standard liquid handling systems). The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described. All documents cited in this specification include, but are not limited to, patents, patent specifications, technical documents, books, and papers, which are hereby incorporated by reference in its entirety for all purposes. In the event of any inconsistency between any incorporated document and any term defined herein, the present specification controls. The present teachings are not intended to be limited to such specific embodiments when the present teachings are described in connection with the specific embodiments. Rather, the teachings include many alternatives, modifications, and equivalents, as will be appreciated by those skilled in the art. SUMMARY OF THE INVENTION [0004] In some embodiments, the disclosure is directed to cracking a solid having an attachment or embedded in a biological material. The lysing reagent solution having the composition has one or more of a detergent, a chelating agent, a reducing agent, and an enzyme. The detergent may be an anionic detergent, an ionic detergent or a combination thereof. The detergent may be N-lauric myosinic acid (NLS, also known as laurel sarcosine sodium (Sarc〇sy 丨 or 5〇心1!11118111'〇71531' (;03-inate)), de-milking Sodium cholate, CTAB, Twelfth y?-D-maltose, 壬 基 _N-methyl melamine, polyethylene glycol p-(1,1,3,3-tetradecylbutyl) -Phenyl ether, sodium dodecyl sulfate (SDS) and combinations thereof, or more. The chelating agent may be ethylene glycol tetraacetate (EGT A ) 099131421 Form No. A0101 Page 4 of 54 1003014903-0 201213805 One or more of ethylenediamine tetraacetate (edta), citric acid and combinations thereof. The reducing agent may be tris(2-carboxyethyl)phosphine (TCEP), dithioerythritol (DTE) and one or more of dithiothreitol (DTT). The enzymes may be caspase, chymotrypsin, pepsin, protease kappa, thrombin, Staphylococcus V8 protease, chain Among them - or more of mold protease, papain, Bacillus sp. E1A protease and trypsin. Salts can be gasification, gasification, gasification, gasification and gasification. One or more of the deuterated forms. This cleavage 0 solution can have a range from 5.0 '12.0, or 5.1, 6.0, 7.0, 8. 0 9. 〇' 1 〇. G smashed 11. 〇 and in every ― The pH in the interval between the integers. In some embodiments, the solid sample that is cleaved by lysis nightly has a nucleic acid. This can be a biological material, an adhesive substrate, or a natural or beta-forming X. In some embodiments, the biological material may be bone, cartilage, band, tendon, and teeth. In some embodiments, the adhesive material may be chewing gum, snow, buttocks, pedicle, film, and adhesive. Tibetans, stickers, paper, smudges, envelopes, envelopes, postage (such as postage stamps) and tape. In some embodiments, 'biological samples can be embedded or adhered to adhesive materials. In many implementations In the example, the skin patch is selected from the group consisting of an electronic electrode, a transferable totem, a skin infiltrated chemical patch, and a wound care dressing, and the adhesive tape can be an adhesive bandage, a tape, a packaging tape, Duct tape, I tape, hair tape, fingerprint tape peeling, and the like. In the specific embodiments, the method disclosed is the use of the disclosed lysing solution 1003014903-0 liquid to produce a product comprising a nucleic acid. The method includes the lysing solution 099131421 Form No. Α0101 Page 5 of 54201213805 A solid sample is cultured in a liquid, and the supernatant of the nucleic acid extracted from the solid sample is extracted. The solid sample may be, for example, bone, cartilage, tendon, and teeth. Materials such as chewing gum, snowy buttocks, finishing pedicles, adhesive films, adhesive labels, stickers, skin patches, envelopes, envelope seals, stamps, peeling of fingerprint tapes, adhesive materials for tapes, denim materials. In some embodiments, the method may also include shaking and/or vortexing the solid sample in the lysis solution when the mixture is cultured, centrifuging the solid sample in the lysis solution after culturing, and lysing the solution in the lysis solution. The biological sample, the biological material, and/or the solid sample are cleaved to generate a lysate having the nucleic acid and extract the lysate to obtain a product carrying the nucleic acid. In other specific embodiments, the disclosed method is a method of isolating nucleic acids from a solid, wherein the solids are cleaved via a lysis solution and the lysate is centrifuged to separate the solids from the nucleic acid still in the supernatant. In some embodiments, the method can also include shaking the solids accompanying the lysis solution and extracting the lysate to obtain a product accompanying the nucleic acid isolated from the solids. In other embodiments, the removal of nucleic acid from the fingerprint tape using the methods described herein can be used to identify an organic material from which the biological material is derived. This nucleic acid can be used for genotyping assays for STR, HLA markers and RFLp cutter i, Υ-STR and Υ-SNP typing, mtDNA sequencing, and insertion/deletion sigmoid typing. Fingerprints can be used in conjunction with genotyping tests to separate fingerprint collection and/or fingerprint databases for identification purposes. Other specific embodiments include biomaterials, adhesive materials or substrates, and denim materials or form numbers for use, for example, from epithelial cells, blood, semen saliva, or other biologically known 099131421 fluids known to those skilled in the art. A0101 Page 6 of 54 1003014903-0 201213805 Kit for extracting nucleic acids from a solid. This kit has one of a first detergent, a chelating agent, a reducing agent, and an enzyme as a separate solution or combination for each. In some embodiments, the kit may also have an optional solution such as a solution having a polymer. The polymer comprises one or more of dextran, cellulose, cellulose derivatives, soluble starch, dextrin, cell dextrins, polyethylene glycol, heparin, glycogen, and combinations thereof. The kit may also have a second cleaning agent, a cleaning solution, a second cleaning solution, water without D N a s e , a magnetic device, and particles that are magnetically attracted to % (magnetic nanoparticles coated with dextran). It will be appreciated that the specific embodiments provided are not required to have all of the aspects and features described herein. It should be understood that these aspects and specific embodiments are merely illustrative and illustrative and not restrictive. The applicant provides a specific implementation as a method of extracting nucleic acids from a sample containing a small amount of nucleic acid. Specific embodiments of such methods are rapid, free from the use of organic solvents, simplified steps, and reduced risk of sample handling and cross-contamination with improved user safety. > These and other features of the present teachings are set forth herein. [Embodiment] The following definitions are used for the purpose of interpreting this specification and, where appropriate, the terms used in the singular will also include the plural and vice versa. When any definitions set forth below conflict with the usage of that word in other documents (including any documents incorporated herein by reference), unless clearly stated to the contrary (eg, in the document originally used), The definitions set forth below are intended to serve the purpose of interpreting this specification and its related claims. It should be noted that, as used in this specification and the appended claims, unless explicitly and unambiguously limited to 099131421 Form No. A0101 Page 7 of 54 Page 301014903-0 201213805 - Refers to the 'singular form' - (a), "an" and "the" include the plurals. Unless otherwise stated, the use of "or" means "and/or". For the purposes of illustration and not limitation, "χ and / or Y" may mean "X" or Γγ" or Γχ" and "γ". Use "comprise", "include (c(10)prises), "comprising", "include", "includes" and "inciuding" as mutually replaceable and non-intentional For the limit. In addition, if the description of one or more specific experimental examples uses the term "comprising", those skilled in the art will appreciate that in some specific examples, this particular embodiment (emb〇di_ raent) or specific embodiment (emb) 〇dim.e.nts) may be alternatively described using a language consisting of "consisting essentially of" and/or "consisting 〇f". The term "and/or" means one or all of the listed elements or any two or more recited elements. The practice of the present invention can be carried out using conventional techniques as well as descriptions of organic chemistry, polymer technology, molecular biology (including recombinant techniques), cell biology, biological robes, and immunology within the skill of the art. Such traditional techniques include nucleoside acid synthesis, hybridization, extension reactions, and the use of markers to detect hybridization. Specific descriptions of suitable techniques can be obtained by reference to the examples described herein below. However, other equivalent traditional degrees can of course be used. Such traditional techniques and descriptions can be found in standard laboratory manuals such as Genome Analysis: A Laboratory Manual Series (Vols. I-IV), PGR Primer: A Laboratory Manual, and Molecular Cloning: A Laboratory Manual (all from Cold Spring) Har- 099131421 Form No. A0101 Page 8 of 54 1003014903-0 201213805 bor Laboratory Press, 1 989 ) 'Gait, w〇ligo-nucleotide Synthesis: A Practical Approach” 1984, IRL Press, London, Nelson and Cox (2000) ),

Lehninger, Principles of Biochemistry 3rdEd., W. H. Freeman Pub., New York N.Y. and Berg et al. (2002) Biochemistry, 5th Ed., W. H. Freeman Pub., New York, N.Y. All of the objects are hereby incorporated by reference in their entirety for all purposes.如同 As used herein, the term "attached" means a transition between at least two objects or surfaces, or at least two objects or surfaces joined together. This bond can be a common fusion, bonding or pasting of at least two objects or surfaces. As used herein, the term "adhesive" means a substance that is capable of binding at least two objects. Adhesive can be cement, glue

Water, paste, powder, glue or chemical compounds or polymers from natural or synthetic sources' can be applied to envelopes, films, labels, paper, patches, stamps or tapes as one of the two objects. The adhesive can be a liquid or solid or can be solidified by evaporation or heat, UV curing or other physical means to dry. The first object may have its indispensable adhesive (e.g., glue) or an adhesive (e.g., tape) to which it is applied. Alternatively, the first object comprises an adhesive to enable the first object to engage at least the second object in a removable, temporary, semi-continuous or continuous state corresponding to the bond formed thereby. The adhesive can be made soluble or viscous via the application of a liquid such as saliva. For example, a stamp is added to the tongue after it is used for sealing. Another example can be an envelope seal 099131421 mouth, which when wiped is made the adhesive on the envelope seal into the form number A0101 page 9 / total page 54 1003014903-0 201213805 eight I〗, and then When folded to close the opening of the envelope, the seal is created as the adhesive maintains a continuous, almost permanent envelope contact of the envelope seal. As used herein, the terms "adhesive material" and "adhesive" are used interchangeably and mean that the object contains or has an adhesive that is indispensable to its composition. Adhesive surface or adhesive composition, such as chewing gum, cigar butt, cigarette butts, adhesive film, adhesive labels, stickers, skin patches, envelope seals, stamps, tapes used to peel fingerprints, biological, organic or inorganic materials And tape. As used herein, the term "adhesive f" means a thin, elastic sheet that can be attached to an object. The film can be attached to an object or can be removed. As used herein, the term "adhesive label" or "a (jhesive sticker)" means an object that has identification information or design: on the upper surface, as opposed to identifying information or design. The surface is attached to the 'other side' or the surface. This label can be attached to other objects or moved in. As used herein, the term "adhesive paper" means the accompanying adhesive surface. Paper (cellulose, wood pulp derived, vinyl, plastic or synthetic paper) that forms a removable, temporary, semi-permanent or permanent bond corresponding to the surface to which it is attached, such as envelopes, stamps, Refillable solid or viscous note or sticker 〇 As used herein, the term "adhes i ve skin patch" means attached to a removable form Integrity or subject to 099131421 Form No. A0101 Page 10 / Total 54 Page 1003014903-0 201213805 Round, oval, rectangular or other shapes of the skin surface of damage (injury) such as EKG electrodes, transferable totems, nicotine patches and plaster As used herein, the term "adhesive tape" means a flexible band or ribbon that can be attached to another object or surface, such as sports tape, packaging tape, Scotch® brand tape. The tape may be permanently attached to other objects or surfaces or may be removable. The tape may be soluble or insoluble in an aqueous or organic solution. As used herein, the term "biological material" means Refers to blood, mucus, semen, saliva, skin tissue, bone, cartilage, ligaments, tendons, teeth, fingerprints, etc. Biomaterials can be biologically derived, hardened, solid, and natural compositions. A solid, fluid, tissue, or cell comprising a nucleic acid. The biological material can include, but is not limited to, a nucleic acid comprising self-adhesive and/or viscous Material to be recycled, including but not limited to chewing gum, cigar butt, cigarette butts, adhesive film, adhesive labels, stickers, skin patches, envelopes, envelope seals, stamps, fingerprint tape peeling and tape As used herein, the term "biological sample" means a biological material that is attached or intended to be incorporated into an adhesive material, such as blood, mucus, semen, saliva, skin cells, skin tissue, bone, cartilage, Ligament, tendon, tooth, fluid, tissue or cell. This biological sample contains nucleic acid. As used herein, "DNA" means various forms of DNA, such as genomic DNA, as is known in the art. , cDNA, isolated nucleic acid molecules, plastid DNA, and chromosomal DNA. "Nucleic acid" means any form of nucleic acid molecule or molecule (molecules 099131421 Form No. A0101 Page 11 of 54 1003014903-0 201213805), DNA or RNA (ribonucleic acid). As used herein, the term "nucleic acid molecule" or "extracted nucleic acid" means a nucleic acid molecule (DNA or RNA of any form) that has been recovered from its natural environment. Some examples of extracted nucleic acid molecules are partially or substantially purified nucleic acid molecules, nucleic acids obtained by forensic and other samples containing biological materials, such as liquid, mucus, semen, saliva, skin tissue, bone, Cartilage, ligaments, tendons and teeth, etc. Also conceived are biomaterials that are self-adhesive and/or viscous material recovery, including but not limited to chewing gum, cigar butt, cigarette butts, adhesive films, adhesive labels, stickers, skin patches, envelopes, stamps and tapes, and Nucleic acid samples from archaeological, criminal field processing, forensic, human identification, natural and large-scale disaster sites collected through forensics, crime, research, search and rescue, and recycling methods and techniques. As used herein, the terms "fingerprint" and "fingerprint image" are used interchangeably herein and refer to the pressure of the ridge of the skin that is created when the ridges and depressions of the skin epidermis are found on the ventral surface of the finger. Trace friction, including but not limited to fingers and toes, thumb and palm and single sole contact surfaces. As used herein, the term "genetic DNA" refers to a nucleic acid comprising a chromosomal DNA sequence of a gene or a fragment of a gene comprising a non-coding region of a DNA sequence and a coding region. The gene D N A also means DNA that is directly isolated from a biological sample (eg, all or part of such a MA of bone, tooth, tissue, cell or chromosome or a copy of vegetative reproduction). Genome MA also refers to DNA extracted from adherent and/or viscous materials such as chewing gum, cigar butt, cigarette butts, adhesive films, adhesive labels, stickers, skin patches, envelopes, stamps and tapes. The gum can be a natural or synthetic source. 099131421 Form No. A0101 Page 12 of 54 1003014903-0 201213805 Natural rubber can be latex, Ala, Gicle and Chicle varieties such as Chico and Rubber Tree (chic〇0 and Chic〇zap〇te). The synthetic gum can be a polyene. As used herein, the terms "rough twill material" and "thick twill" are used interchangeably and refer to materials of any animal or plant origin. The denim material is completely blended with cotton or cotton natural or. Made up of fiber. Cotton denim fabric may include, but is not limited to, blue

Dark blue, black, stone wash, pickled, and the like. As used herein, the terms "polynucleotide", "oligonucleotide" and "nucleic acid" are used interchangeably herein, and mean single-stranded and double-stranded polymers of nucleomonomers, She does: not about 2,-deoxyribonucleic acid (DNA) and ribonucleic acid (RNA) 'its its interaction via nucleotides of phosphoric acid/nucleic acid (4) and related counterions (eg 11 NH4, II) Connected to the base, Mg2+, Na+, etc.). Polynucleic acid can be completely depleted by ribonucleoside (IV), h-ribose or its fungus

s constitutes ' and can include analogs of nucleotides. The nuclear acid monomer unit may comprise any core (4) or nuclear hearing analog. The size of the poly(tetra) acid type is from a number of monomeric units (such as 5-40 (although they are sometimes referred to in the art) to thousands of monomeric cores (quad). Unless otherwise indicated, 'each time indicates a multicore. In the list, it will be understood that the nuclear acid is from 5 to 3 in order from left to right, and "A" represents deoxyadenosine, "C" represents deoxyacid, and "G" represents Oxygenic acid, "τ" stands for deoxygenated_« and "U" stands for deoxycortic acid, unless otherwise indicated. 099131421 Page 13 of 54 This article is used in this article, the term "solid" means the facet The material form number Α0101 « ι〇--1003014903-0 201213805 materials, such as adhesives, which contain materials and hardened, dense materials with matrix, such as maternal biological materials such as: bone, teeth, and cartilage, Tendons and ligaments and common powders composed of exfoliated skin cells. This solid may contain or migrate to contain nucleic acids. As used herein, the term "solid sample" means a hard, dense material, possibly a biological material, Such as bone, cartilage, ribbon, tendon Adhesive material with or without pores and/or insoluble in water, and containing or suspected of containing nucleic acids. At the same time, the enumeration of numerical ranges and the numerical ranges on the endpoints include all numbers that are included within the range (eg 1 to 5 include 1, 1.5, 2, 2. 75, 3, 3. 80, 4, 5, etc.) In some embodiments of the present teachings, in the description of the compositions, methods and kits The nucleic acid molecules can be isolated and/or extracted from the sample, and in some embodiments, the products made by the compositions, methods, and kits are nucleic acids. In some embodiments, the compositions of the present teachings , methods and kits result in the production of a product comprising the extracted nucleic acid. Compositions are provided in some embodiments of the present teachings, wherein the sample may be accompanied by at least one detergent, chelating agent, reducing agent and enzyme, and combinations thereof. Each lysis solution is treated to extract nucleic acid molecules from the sample. In various embodiments, the sample may comprise one or more free nucleic acids; extraction from the solid substrate includes, but is not limited to, organisms Materials such as bone, cartilage, ligaments, tendons and teeth, etc. In various embodiments, the sample may comprise nucleic acids extracted from an adhesive material including, but not limited to, for example, chewing gum, cigar butt, butt, adhesive film, Adhesive labels, stickers, skin patches, envelope seals, stamps, fingerprint tape peeling and tape materials. In some embodiments, the sample can be 099131421 Form No. A0101 Page 14 / Total 54 Page 1003014903-0 201213805 Contains The nucleic acid extracted from the twill material, Α, 匕, but not limited to blue, dark blue, black, stone washed, pickled to cover such a long uncle of the cloth. In this teaching, the additional implementation is described in this article. A method by which an acid can be extracted from a sample, comprising the step of accommodating the sample with a nuclear multi-cleaner, a chelating agent, a reducing agent, a salt and an enzyme, or extracting a supernatant. Cultivation can be in the room w. Raise 5 C, j π C, 41 ° C, 42 ° C, 43 ° C, 44-C, 45. (:, 5frr 〇u L, 51.c, 52〇C ' 53〇C ' 54〇C ' 55〇C ' 56〇C ^ 57°r 7 C, 5»°C, 59 Ο t town, heart and WC, and the interval therein. This method may further comprise the steps of removing the solid from the cultured solution and removing the nucleic acid from the supernatant, and the step of treating the supernatant to separate the nucleic acid &

The nucleic acid thus obtained can then be used in any of a variety of downstream applications and assays, such as, for example, quantification, detection and genotyping of specific nucleic acids or even biological species. For example, these analyses can be performed via pcR amplification. As an example, in forensic DNA analysis, human nuclear DNA (nDNA) and/or genomic DNA can be obtained from complex biological materials and then genotyped using PCR. As another example, qNa preparation can be used to quantify human DNA (or more specifically, male human DNA) using, for example, the Quantifiler® or Quantifiler Duo® kit (Ap_plied Bi〇systems/Life Technologies C〇rp Foster City, The instant polymerase chain reaction system of CA), and/or, for example, uses a system such as the AmpFISTR® kit to genotype a short tandem repeat site for a somatic chromosome or a Y chromosome. Depending on the amount of DNA present in the sample, a particular genotyping system can be selected to generate the best results for a particular analysis. Therefore, in order to optimally use nucleic acids in downstream applications, extraction and separation methods are particularly required. 099131421 Form No. A0101 1003014903-0 Page 15 / Total 54 Pages 201213805 Only results in the production of the same cover, but also relatively no downstream applications The product of an inhibitor (e.g., PCR). Nucleic acids that are analyzed by quantification, detection, and genotyping can be used to determine the source of the nucleic acid. In a typical place, forensic evidence samples are subject to change in the type, substance or matrix of the sample, wherein the biological material is embedded, including but not limited to, solid biological samples or surfaces of forensic samples collected, It may cause damage to the sample from exposure to the environment, contamination by the PCR inhibitor, and restriction of the starting sample material. Nucleic acid extractions, including but not limited to forensic evidence samples, which are often exposed to changes in temperature, excessive humidity or dehydration, and sample perspiration as known to those skilled in the art in the course of acquisition. It may lead to difficulties in isolating tannic acid as well as potential compound sweating's, for example: other chemical treatments or dyes that act to suppress the denier of pCR, and thus attempt to interfere with genotyping or other assays. A modified extraction procedure is required, and in doing so, the inhibitor is removed during the isolation of the nucleic acid, such as DNA or RNA for forensic analysis (for example, an enzyme that is amplified prior to use in subsequent steps). As an example, a forensic sample can be used as an adhesive material or as part of the sample itself. Adhesion, the presence of ruthenium often reduces the overall recovery of nucleic acids, as the separation of nucleic acid adhesives using granules may interfere with the separation step. In an example exemplified below, nucleic acids extracted from cigarette butts, chewing gum, and dried blood, saliva, and biological materials (including skin cells and cells from fingerprints) in tape stripping result in higher nucleic acid recovery and when The short tandem repeat (str) analysis is depicted as a no pCR contaminant when depicted in the graph. Another example of a difficult forensic sample is a solid biomaterial including, but not limited to, calcified tissue such as bone and teeth. This dense substrate needs 1003014903-0 099131421 Form No. A0101 Page 6 / Total ^ Page 201213805

The lengthy, labor-intensive process often lasts for more than 24 hours before the trace amount of citric acid is extracted. The teachings of the present disclosure achieve maximum amount of DNA recovery by disrupting the calcification matrix while contacting most of the biological material with the lysis reagent solution. The yield of 1) questions from bone and tooth samples is summarized in Table 1. In the example, note the time required to extract the nucleic acid from the bone and teeth to achieve the desired yield only after two hours. Because the disclosed lysing solution composition and the method of extracting nucleic acid result in improved and faster DNA recovery, the amount of DNA present in the sample can be more sensitively detected by an instant polymerase chain lock known to those skilled in the art. Detection by the method of reaction. Various embodiments of the present teachings relate to efficient extraction of nucleic acids, such as, for example, the amount of genomic DNA such that: the Prep-FilerTM forensic extraction kit is used (P/N 4392852 or 4392353,

The subsequent isolation and elution of the nucleic acid by Applied Biosystems, Foster City, CA provides nucleic acids suitable for quantification and analysis of DNA. Thus, specific embodiments of these teachings enable the efficient extraction of nucleic acids (e.g., genomic DNA) from various solid species of biological materials, materials containing adhesives, and biological fluid samples from denim. In addition, nucleic acids such as genomic DNA can be isolated from small amounts or large amounts of biological material that is typically processed in the laboratory, such as, for example, those involved in genotyping. Specific examples of the lysis solution provided herein may comprise a mixture of, for example, tetraacetic acid glycol (EGTA) and/or ethylene tetraacetate (EDTA) and/or citric acid chelating agents (preferably 250-500). mM is a range); for example, at least one of N-lauric acid, sodium deoxycholate, CTAB, N-dodecylate; 5-D-maltosine, thiol-N-methyl branamide, Triton ® X-100, NP-40 and/or sodium lauryl sulfate detergent (preferably 099131421 Form No. A0101 Page 17 of 54 page 1003014903-0 201213805 with a range of 0.1% to 3%) Such as sodium chloride, potassium chloride, magnesium chloride, manganeseated gas and salts thereof in the form of fluorinated and iodinated (in the range of 50 mM to 500 mM); and, for example, tris(2-carboxyethyl)phosphine ( TCEP), a reducing agent of dithioerythritol (DTE) and dithiothreitol (DTT) (preferably in the range of 10 to 100 mM), and, for example, at least one caspase, pancreas Chymotrypsin, pepsin, proteinase K, thrombin, Staphylococcus V 8 protease, pronase, papain, Bacillus sp. p. E1A protease and trypsin Hormone (preferably 0.5 to 1 mg / mL of the range). The table provided below indicates the extraction efficiency as a comparison of the various incubation times of the lysis solution, the amount and type of starting solids, the adhesion or the denim material. The lysis solution extracts the nucleic acid from the solid substrate or adhesive material so that the nucleic acid supernatant can be separated and the impurities are washed using the Pr epF i i er711 kit. These specific embodiments and other features of the present teachings will become more apparent from the description herein. Various embodiments of the present teachings relate to a lysis solution and a method of using such a solution (which can be assembled into a kit by inspection) for use in containing biological samples and, for example, biological samples (considered to contain or have been The adhesive material of the solid sample containing the nucleic acid is extracted, and the nucleic acid includes, but is not limited to, genomic DNA. Adhesive materials can be stripped, for example, by forensic methods, such as dry blood, saliva, and fingerprints, as well as envelope closures, stamps including, but not limited to, stamps, coupons, hunting license stamps, protective stamps, chewing gum. , cigar butt, cigarette butts, adhesive films, adhesive labels, stickers, skin patches, envelopes, stamps and tapes, and more. Adhesive materials are considered to contain nucleic acids attached or embedded in an adhesive material, and the teachings are related to the extraction of nucleic acids from a sample of adhesive material 099131421 Form No. A0101 Page 18 of 54 1003014903-0 201213805, if any . Adhesive materials have been shown to interfere with subsequent nucleic acid isolation and overall yield when using the Bulk-FilerTM kit. Adhesives interfere with the operation of magnetic particles in the polymer-nucleic acid to magnetic Z particle composites produced by the prepFiler. The formation of this complex is such that the polymer traps the nucleic acid, the polymer attaches to the magnetic particles and thus indirectly binds the magnetic particles to the nucleic acid until the complex is washed to remove contaminants and inhibitors such that the nucleic acid is correctable For downstream applications, such as PCR, obtaining nucleic acids from an adhesive containing a sample is difficult, cumbersome, and low-yielding prior to the development of the lysis solution taught herein. Dyes, pigments, and fibers found in denim fabrics (including, but not limited to, blue jeans, skirts, and jackets) used in manufacturing inhibit the STR analysis of the nucleic acid isolated from the genotyping test in the genotyping test' Iden-tifiler® kit (Applied Biosystems). The fibers appear to complex with pigments or dyes and interfere with the operation of the magnetic particles in the polymer-nucleic acid-magnetic particle composites produced by the PrepFiler program. This results in damage to the nucleic acid obtained from the denim sample by PCR inhibitors, which are difficult, cumbersome, and low-yielding prior to the development of the Tris-Cl lysis solution taught herein. Various embodiments of the present teachings relate to, for example, nucleic acid extraction for genomic DNA, including lysis solutions, methods and kits for extracting nucleic acids from solid biological samples and adhesives, including Not limited to 'bone, cartilage, ligaments, tendons and teeth. Specific embodiments of these methods can include: cleaning the solid sample with a mild detergent solution, air drying overnight, removing the outer layer with a matte stone, and smashing the sample with the mound. The prepared sample is then incubated with the lysis solution to release the nucleic acid containing cells 099131421 Form No. A0101 Page 19 of 54 1003014903-0 201213805 from the solid substrate. This solid biological material contains nucleic acids, and the teachings are related to the extraction of nucleic acids from solid materials. Calcified and hardened biomaterials have been shown to provide low yield nucleic acids when using the PrepFilerTM kit (Appl ied Biosystems, Foster City, CA) because solid matrices offer particular challenges for nucleic acid extraction and separation. Nucleic acid production from solid biomaterials has been improved prior to the PrepFiler program using the lysis solution taught herein. Standard nucleic acid isolation techniques, including (cell lysis and washing and rinsing of nucleic acids), are well known in the art and, unless otherwise indicated, may be described by various techniques, for example, by describing A Typing Protocols: Molecular B ίοlogy and Forensic Analysis, lstedition, B. Budowleet ai..eds., Eaton Publishing Co. (2000) ; JM But 1er, Forensic

DNA Typing: Biology, Technology, and Genetics of STR Markers, 2ndedition, Elsevier Aca-demic Press (2005); or P. Giil, "Application of Low Copy Number DNA Profiling," Croatian Medical Journal42 : 229^-232 ( 2001 ) ; FR

Bieberet ai,, "Isolation of DNA from Forensic Evidence,"Current Protocols in H uman G ene t- ICS, Supplement 26 (2000) ; Forensic DNA Pro filing Protocols, Metiiucls in Molecular Biology, vo1. 9 8, PJ Lincoln and J. Thomson, eds., Humana Press (1998) ° Applicants have found that when using PrepFiler's plan for adhesive materials containing solid nucleic acids and solid biological samples, use the text 099131421 Form No. A0101 No. 20 Page / Total 54 pages 1003014903-0 201213805 A lysing reagent solution with one or more detergents, chelating agents, reducing agents, salts and enzymes, nucleic acids can be more easily isolated and purified. The lysis solution improves the yield and efficiency of nucleic acids (e.g., genomic DNA) from adhesive materials, solid biological samples. The detergent can be polar or non-polar. Some examples of suitable cleansers include, but are not limited to, N-lauric myosinic acid (NLS), lauric acid sarcosine (also known as sar-cosyl), a derivative derived from sarcosine. Ionic surfactant; hexadecyltr i methyl ammonium bromide or cetyltrimethylammonium bromide (CTAB); deoxycholic acid; sodium citrate; sodium deoxycholate; twelfth base / 3-D- Maltose; thiol-N-methyl branamide; sodium lauryl sulfate; polyethylene glycol p-(l,1,3,3-tetramethylbutyl)-phenyl ether (generally called For Triton® X-100); and combinations thereof. In some embodiments, the cleaning agent comprises NLS in the range of 0.5 to 3% w/v. Some examples of suitable chelating agents include, but are not limited to, ethylene glycol diamine (EDTA), ethylene glycol bis(2-aminoethyl)-hydrazine, hydrazine, hydrazine, hydrazine in the range of about 250 to 750 mM. '-Tetraacetic acid (EGTA) with citric acid. Some examples of suitable reducing agents include, but are not limited to, tris(2-carboxyethyl)phosphine (TCEP), dithioerythritol (DTE), and dithiothreitol (DTT) in the range of about 10 to 100 mM. (Freshly prepared before use). Some examples of suitable enzymes include, but are not limited to, caspase, chymotrypsin, pepsin, proteinase K, coagulase, staphylococcus in the final concentration range of 0.5 to 10 mg/ml in the lysis solution. V8 protease, pronase, papain, Bacillus sp. E1A protease, and trypsin. In some embodiments, the lysis solution is combined with a chelating agent at a pH of about 7.5 to 8.5, followed by a detergent and a salt, followed by a reducing agent solution and an enzyme. 099131421 Form No. A0101 No. 21 Page / Total 54 pages 1003014903-0 201213805 Preparation. In some embodiments, the chelating agent is prepared in a solution with a cleaning agent, with or without a salt, the reducing agent is freshly prepared, and the enzyme is in a 'liquid. Salts are in solution or present in detergent solutions. The Lok solution can then be combined to form a lysis solution prior to use. Alternatively, in some embodiments of the present teachings, the lysis solution can be prepared and then dried. λ; Donggan's lysis reagent can be dissolved before use. The method of such bundle drying is conventional and is known in the art as a result of the freshly prepared cleavage solution known to those skilled in the art. The result is that the component is not degraded or oxime, for example, a cause may exist in long-term storage or exposure. Total yield under light and thus reduced acid extraction and nucleic acid. In some embodiments, the adhesive material or solid biological material is cultured for about two hours from about to the next to release the cells from the adhesive material. The solid biological matrix is 56 〇c which is a commonly used temperature. In some embodiments, the natural or synthetic material is from about 4 G mo. (: culture for 40 knives to 1 hour. This extraction can be assisted by shaking. In some embodiments, the lysis solution further comprises a compound that protects the released nucleic acid and maintains their compatibility in downstream assays, for example, by way of example For the PCR test and in the specific DNA genotyping system, for example, the str test. After culturing the adhesive material or the solid biological sample, the sample can be centrifuged to granulate residual solid tissue or adhesive material. The resulting supernatant product comprises a nucleic acid. This supernatant is suitable for use in nucleic acid isolation and purification (using methods known to those skilled in the art or commercial core 1 as a separation product, such as pr epF i 1 er TM kit). This kit can be used to remove cell debris, residual contaminants and/or pCR inhibition 099131421 Form No. A0101 Page 22 / Total 54 Page 1003014903-0 201213805 In this tutorial In some embodiments, the nucleic acid molecule can then be separated from the solid matrix 'including but not limited to: bone; cartilage; kinetic band•' tendon; and teeth. In some embodiments, the biological sample is calcified, dried, dried, or otherwise in a natural or preserved state. In some embodiments, the 'solid biological sample is dispersed, comminuted, ground, or otherwise ground. Fine particles (eg powder). The yield of a nucleic acid (eg DN a ) varies depending on the age of the solid material, the amount of starting material and the q degradation or storage of the sample. The production of nucleic acids (eg DNA) from bone and tooth samples Summarized in Table 1 MA MA yield range from 100 mg long bone samples is from (L25 to 2.15 ng 'which is typical for such sample types because each bone sample is unique in the inclusion and state of the biological sample The values for the internal PCR control group (IPC) were determined by the Quantifiler® Human DNA Quantitative Kit (Applied Biosystems) compared to the value of the no template control group (NTC, Table 1). There was no significant increase in the bone sample q tested. If the DNA extract contains a PCR inhibitor, typically a sample is expected to be compared to the IPC of the NTC (the value is shifted above (Applied Biosystems, Quantif) Iler® Human DNA Quantitation Kit and Quantifiler® Y Male Human DNA Quantitative Kit. User Manual. PN 4344790 Rev. B, (2003)). Therefore, PCR inhibitors present in bone and tooth samples during DNA extraction It has been efficiently removed (using a lysis reagent solution followed by separation and purification with the PrepFilerTM kit reagent) 099131421 Form No. A0101 Page 23 of 54 1003014903-0 201213805 Table 1: Human DNA production from bone and teeth Sample a describes total yield (ng) IPCCT number tibia-1 100 mg long bone 1.9 27.05 bone-2 100 mg long bone 0.66 27.12 bone-3 100 mg long bone 1.19 26.96 bone-4 100 mg long bone 1.6 27.19 bone-5 100 mg long bone 0.8 27.12 bone*6 100 mg long bone 0.24 27.11 bone-7 100 mg long bone 2.22 27.17 bone-CA 50 mg clavicle 7.95 ±1.2 27.012 tooth 10 mg 450 ± 50 26.95 extraction blank group 0 26.80 no group ( NTC) The age of the NAb NAb a bone sample is unknown. The tooth sample is 1 year old. b Not applicable for the optimal extraction and yield of nucleic acids. The amount of bone sample varies from 50 mg to 200 mg, and whenever necessary, the volume of the supernatant of the lysing solution collected is separated in the DNA using PrupFilerTM. The buffer was adjusted to 200 μm. When the amount of bone powder was increased from 25 mg to 50 mg of powder, the total yield of DNA was almost double (Table 2). Further increase in the amount of bone powder increases DNA yield, but the amount of DNA production increase is not proportional to the amount of bone powder. This evaluation was repeated in triplicate. After culturing 200 mg of bone meal, the recovered lysis solution supernatant volume was 130 μί (compared to 200 μm from 50 mg bone powder). Therefore, treatment of more than 200 mg of bone meal did not significantly increase DNA production. 099131421 Form No. A0101 Page 24 of 54 1003014903-0 201213805 Table 2: Bone-CA (mg) Total Yield (ng) IPCCt per gram of bone 値25 4.8 ±1 0.2 27.05 50 9.3 ±3 0.18 26.94 100 12.0 ± 3 0.12 27.03 200 21.6 ±6 0.11 26.89 Extraction blank group 0 NA NA Moldless chick group (NTC) NA NA 26.85 The degree of extraction from the bone sample nucleic acid was investigated at different time intervals from 1 to 18 hours. 50 mg of bone was extracted in triplicate. DNA yields are summarized in Table 3. When the extraction time of the bone sample with the lysis solution was increased from 1 hour to 2 hours, the DNA yield was increased by 1.54 times. However, further increases in culture time to 18 hours did not substantially increase DNA production. However, up to 18 hours of incubation time has no adverse effects on the extraction. Table 3: Lysis time (hours) Total yield (ng) IPCCt number 値1 5.2 ± 1 27.132 2 8.0 ± 1 27.047 4 8.5 ± 1.5 26,952 8 8.8 ±1.5 26.913 18 11.4+1 26.923 Faceless control group NA 26.815 by manufacturer Project book, using AmpFlstr® 099131421 for STR analysis Form No. A0101 Page 25 of 54 1003014903-0 201213805

Identifiler® and MiniFilerTM kits (Applied Bi〇Sys — terns) were used for DNA extraction of seven long bone samples (bone 1 to bone 7). The results are summarized in Table 4. This I d e n t i f i 1 e r kit amplifies all 16 STR loci' and the MiniFiler kit amplifies 9 STR loci. In each test sample, the final locus is compared to the total expandable locus for each set of groups. The Bone-4 sample provides a complete STR profile (using two sets). Bone-6 did not provide results when amplified using the Identifiler kit, but the MiniFilerTM kit provided amplification of only 4 sites. All other bone samples were tested in a M i n i F i 1 erTM kit to provide a complete STR profile and a partial STR profile was provided with the Identifiler® kit test. This MiniFilerTM kit is designed to obtain contours from degraded DNA samples (J.J. Mulero, et al., J.

Forensic Sci. (2008) 53:838-852). This result indicates that the bone sample is aged and degraded to varying degrees. Table 4: Sample 3 Iden邯ler® Set MiniFilerTM Set Bone-1 9/16 9/9 Bone-2 9/16 9/9 Bone-3 10/16 9/9 Bone-4 16/16 9/ 9 眚-5 9/16 9/9 Bone-6 0/16 4/9 Bone-7 15/16 9/9 The age of the bone sample is unknown. 099131421 Form nickname A0101 Page 26 of 54 1003014903-0 201213805 In the specific embodiments, the present teachings include the use of multiple samples ranging from the presence of trace amounts of biological material. The sample size can range from 5 in each tube. Feeding tissue or solid biological samples up to 20 ,, then combined with the supernatant and concentrated by standard methods prior to isolation and purification. In the present teachings, the nucleic acid molecules may be separated by an adhesive material, including but not limited to: sigma, snow, buttocks, cigarette butts, adhesive films, adhesive labels, stickers, skin patches. , with seals, stamps and dried blood stripped by tape, dry saliva, including but not limited to dust, earthworms, environmental sampling of dirt and tape. In some specific implementations, skin-like materials include, but are not limited to, EEG or EKG electronic electrodes; transferable genus; skin-permeable chemical patches and wound care dressings. In some embodiments, the tape includes, but is not limited to: tape; sports tape; forensic tape;

099131421. In some embodiments, the sticker includes, but is not limited to, an envelope, a re-adhesive stamp, and the like. DNA from samples containing adhesives (eg, terminal syrup, chewing gum, tape stripping, envelopes, and stamps) using the disclosed lysing solution, PrepFile H buffer, and EZ1 DNA Investigator kit (Qia scn, Valencia, CA) Comparison of production _ 早乂. Not in Table 5. In general, the throughput of the disclosed lysis reagents for the ribbon stripping and the pedicle sample was higher than that obtained with the PrepFiler buffer. Similarly, the DNA production of two of the three test chewing gum samples of the disclosed lysing solution was higher. The sample is usually a similar amount of biomaterial (see Examples I and Iv) 'Because this assessment involves the use of real-world sample types that may be encountered in forensic laboratories, sample species form number A0101 Page 27 of 54 pages 1003014903-0 201213805 Not completely unified. Therefore, this DNA production yield is used to measure the overall performance of the lysis reagent solution. It has been observed that when using PrepFiler lysis buffer, each tape stripping and chewing gum sample releases a large amount of adhesive. For tape stripping samples, some of the adhesive agglomerates can be removed prior to further PrepFiler program processing, however, in many of these samples, magnetic particles cannot form pellets, and some such samples must be Centrifuge during the washing step in the PrepF i er program. The lysis reagent solution has a smaller amount of adhesive released during the extraction process. Therefore, the magnetic particles form stronger pellets during the subsequent PrepFiler washing step, which results in an improvement in nucleic acid recovery including (DNA yield). Small sheets of paper were removed from the pedicle sample by centrifugation. The Sherman brand's final roll paper was purple and the purple dye release was observed under PrepFilerTM lysis conditions. In addition, the IPC CT values for all study case type samples were similar to those for NTC, indicating effective removal of PCR inhibitors from biological samples and substrates (Table 5). 099131421 Form No. A0101 Page 28 of 54 1003014903-0 201213805 Ο

Table 5: STR porch sample surface reagent solution from long bone sample 81 PrepFilerTM mmm Total DNA production (ng) a IPCOr DNA total production® (ng) IPCCt tape stripping with +2pL blood 0 40 26.6 14 27.4 Tape stripping 2 μι·血20 27.3 17 27 Tape + 5 (JL saliva b 143 27.1 64 26.8 5 5 UUl solution 7 26.9 3 27.4 Tape peeling, shirt cuffs 0.6 26,6 NDC ND Tape brake, lining 4 26 , 7 ND ND tape stripping, shirt collar 2.5 26.8 ND ND tape stripping > Shirt collar 0.6 26.9 ND ND Acceptance blank group 0 26.8 0 26.8 Extraction blank group 0 27.0 0 27.0 Winterfresh 7 27.0 15 26.4 Doublemint 6 27.1 3 27.3 Orbit 20 27.1 19 26.6 Extra White 27 28.1 ND ND Airwave (chewing 2 minutes) 7 27.8 ND ND Airwave (chewing 40 minutes) 30 27.7 ND ND Blast Mentos Berry-Lime 30 27.7 ND ND Extract®^White Group 0 27.1 0 27.1 Chedi Marlboro 174 26.7 25 27.6 Marlboro Menthol Light 75 26.7 39 27.1 Sherman 424 26.6 111 29,0 Gauloises Menthol 76 26.7 22 28.0 Marlboro Menthol Light® 35 26.7 3 26.8 Holiday Gold 12 26'9 ND ND Holiday Menthol 37 26.5 ND ND Acceptance blank group 0 26.7 0 26J Extraction blank group 0 26.7 0 26.7 Body sandalwood is b〇uL. The amount presented is the average 値.. <= blood or 5 The team's saliva is directly added to the 1.5 coffee x 2.5 cm tape segment. Wo determines the fragment of the chewing gum 'except Blast Mentos Berry_Lime' are Wrig|ey*s brand · before the analysis 'will be outside the sample _ 72 4 m. This 72 / j view of the face is 4.15X. In some embodiments of the present teachings, a Tris_cl lysis solution can be used to separate a nucleic acid molecule from a denim material that is included on a surface or embedded in a biological flow sample, including but not limited to the 099131421 form. No. A0101 Page 29 of 54 1003014903-0 201213805 Blood, tissue, urine, saliva, for example, hair manure °. Essence (for example, skin cheek cells, vaginal cells, skin, included in fingerprint t) > Amniotic fluid in the cells of the disk and the cells containing the fetal cells, oral samples, packets. A. The odor of the body is moist, wet, dry or amniotic fluid) and semen ° " .. In these specific embodiments, the biological flow attached to the denim material fg body sample may be a sample found in the environment (eg, 4 soil h in other embodiments, (4) exposed to simultaneously lyse the fine material in the biological fluid material H. The method of extracting biological fluid material and extracting nucleic acid from the matrix to extract nucleic acid from the rough twill material. In some cases, the method may also include heating or cultivating the denim with the biological flow material. Quality

Tris-Cl lysis solution. In some embodiments t, the Tris-Cl lysate solution is centrifuged '[beta] to separate the lysate comprising the nucleic acid from the substrate. In some embodiments of the present teachings, compositions are provided wherein the sample may be Tris_cl comprising Tris-Cl, at least one detergent, a chelating agent, a reducing agent, and each of the enzymes and combinations thereof. The lysis solution is treated to extract nucleic acids from the denim substrate. The cleaning agent may be - or more N-lauric sulphate, sodium deoxycholate, CTAB, dodecyl-d maltose, sputum _N_methyl glutamine, d, 3- Tetramethylbutyl stupyl ether

Polyethylene II is intoxicated with P_U sodium sulphate (SDS), and combinations thereof. The chelating agent may be one or more of ethylene glycol tetraacetate (EGTA) and ethylene tetraacetate (EDTA), citrate, and combinations thereof. The reducing agent may be: phosphine (TCEP) disulfide, glycerol (DTE), and dithiothreitol Xueguang DTT). The enzyme can be called "Spicy Sugar (Enzyme, Pepsin, Egg (4) κ Γ 酶, Enzymatic Milk Protein, Pronase, Papaya... Thrombin, Staphylococcus V8 Protease 099131421 Form Number Leg. 1 $ 30 Κ / Total Bacillus licheniformis SP. EU protease and pancreatic egg 1〇〇3〇149〇3~| 201213805 White enzyme. Salts can be one or more of sodium chloride, potassium chloride, magnesium chloride, manganeseated manganese and its fluorinated And iodinated forms. The cleavage solution can have a pH ranging from 5.0 - 12.0, or 5.1, 6.0, 7.0, 8.0, 9.0, 10. 0 and 11.0, and in the interval of each integer. In some embodiments The sample may comprise nucleic acids extracted from denim material, including but not limited to, denim fabrics including blue, dark blue, black, stone wash, pickling, and the like. Additional teachings in the present teachings. In a specific embodiment, the method is described in a method in which a nucleic acid can be extracted from a sample, comprising the steps of: including a Tris-C1, one or more detergents, a mixture of agents, a reducing agent, a salt, and an enzyme. Solution culture; Take the supernatant. The culture can be at room temperature 35 ° C, 40 ° C, 41 ° C, 42 ° C, 43 ° C, 44 ° C, 45 ° C, 50 ° C, 51 ° C '52 ° C , 53 ° C, 54 ° C, 55 eC, 56 ° C, 57 ° C, 58 ° C, 59 ° C, 60 ° C, 65 ° C and 70 ° C, and within the interval. This method may further include the following Step: Centrifuging the culture solution to remove solids from the supernatant, and treating the supernatant to separate nucleic acids. In some embodiments, the PCR inhibitor can be extracted or released to the lysate during the nucleic acid extraction step Or in the extract, this results in poor quality DNA. The quality of the nucleic acid extracted or recovered from the biological fluid sample may be, for example, a nucleic acid analysis step for quantifying, detecting, and genotyping a particular nucleic acid or species.至 U U U U U U U U U U U U U U U U U U U U U U U U U U U U U U U U U U U U U U U U U U U U U U U U U U U U U U U U U Up to 5.0%, and the presence of a chelating agent may be from about 1.0 mM to 600 mM at a pH of 7.0 to 8.5 (pH adjusted with 0.5 -1 M NaOH). Some examples include, but are not limited to, 099131421 Form No. A0101 Page 31 / Total 54 Page 1003014903-0 201213805 'Sodium chloride, potassium chloride, magnesium sulphate, chlorinated and its fluorine in the range of about 1 mM to 1 Μ And chelated forms. In some embodiments, the enzyme concentration is in the range of 1 mg/uL to 50 mg/mL and the reducing agent concentration is in the range of 10 mM to 1 Torr. Use thick twill fabrics of different kinds, weights and colors (from j〇_Ann’ s

Fabrics, San Mateo, CA)) Analysis of the Tris-Cl lysis reagent solution. Basically, blood is applied to each denim type and a 5 mm denim punch is removed from the center of the blood spot (about 1 ^ dry jk solution). Compare Intercolor Balance (ICB) with the Identifi ler® kit (Appl i ed Biosystems) in the denim sample sample listed in Table β, some of which contain the blue dye, pickling treatment, stone disclosed in Figure 1. Wash, ice wash, black dye, texture, print, and different weights and finishes, followed by the accompanying Tris-C Bu Buffer Lysis Reagent Solution, PrepFi lerTM Lysis Buffer and identifi ler® Kit (Ap_ plied) Biosystems) nuclear relatives. In general, the NA quality of the disclosed Tris-C Buffer Cleavage Reagents is sufficient to obtain a complete STR profile for each test sample. These results show that even in the presence of indigo, black stains and chemical treatments (known PCR inhibitors), the disclosed T ris - C Bu buffer lysate solution provides high quality nucleic acids 0 099131421 Form No. A0101 Page 32 / Total 54 pages 1003014903-0 201213805 Form 6: A denim ID Description Old grain description 10OZ Distressed STR 18 70Z blue STR pipe filling -2_ 10.5OZDK Blue stovetop 19 loozsw washed thick oblique tote — 5.80Z not Bleached denim 20 50Z blue fine grain twill 4 90ZDK blue tobacco pipe pants 21 90Z blue texture st ____5 8 0Z blue 隹 pipe pants D 22 90Zmed blue str thick oblique hem 6 ^ 40ZDK blue denim 24 6.50Z 靛 blue denim 7 80Z0K blue denim 25 70Z It blue bamboo thick oblique hemp - 8 pants 27 sbl washed (wsh) thick oblique cloth 1002 —-1_ 9,9〇z m&fm st 28 28. _ 10 11OZ black STR Rough twill 29 10OZ distressed nost _ 11 5 0Z blue twill (XhatGh) denim 30 12.50Z «* str _ 12 12, L BL ice wash 31 40ZII blue twill ~~ 13 β OZ twill (xhatch) denim 32 110Zlt blue brewing _ —Jl_ 10 OZ 靛 blue washed denim 34 4 〇 Zlt blue denim - 15 1002LT blue STRsto 36 70Z blue denim 16 10.5OZmed blue twill (xhatch) 37 90Z light blue wash _ 17 9.750ZU blue stove 38 70Z Color sfr pipe

In some embodiments of the present teachings, the nucleic acid molecules can then be separated from the denim using a Tris-Cl-buffered lysis reagent solution.

Capillary electrophoresis electron micrographs of STR profiles obtained from bones, descent, and some materials containing adhesive swords are typical of such samples. Furthermore, the STR profile confirmed that the DNA obtained did not have any PCR inhibitors (data not shown in 'Stray et al., Forensic Science International: Genetics Supplement Series 2 (2009) - 160' incorporated herein by reference). This lysis reagent solution is shown to disrupt the calcified tissue matrix for efficient DNA extraction from comminuted bone and tooth samples as well as from adhesive materials such as knee bands, chewing gum, envelopes, stamps and final pedicles. As shown in Figures 1 to 4, the results of the STR profile of the denim material confirmed that the obtained DNA did not have any PCR inhibitor. The Tris-C Bu buffered lysis reagent solution was shown to achieve efficient DNA extraction from denim-like 099131421. Form No. A0101 Lysis Reagent Solution and Tris-Cl-Buffer Cleavage Page 33 of 54 1003014903-0 201213805 Reagent solutions all lead to improved nucleic acid recovery, improved DNA isolation, purification and no DNA hazard or hinder further analysis (eg PCR) , Instant polymerase chain reaction and STR profile) interfere with materials and impurities. In other specific embodiments of the present teachings, nucleic acids recovered from fingerprints obtained using the methods described herein as discussed above can be used to identify organisms from which biological materials or biological samples comprising nucleic acids are obtained. ). Fingerprint images can be combined or isolated from nucleic acid identification assays for comparison with forensic databases, including but not limited to fingerprint collection and/or fingerprint databases. Fingerprints are known forensic identification tools, and there are several fingerprint databases and collections around the world for identification and identification/validation purposes. The use of advantages in obtaining nucleic acids from fingerprints provides further confirmation and definitive identification evidence, and further links the identified individuals to appearances and locations or objects that acquire fingerprint nucleic acids. In other specific embodiments of the present teachings, nucleic acids recovered from denim obtained using the methods described herein can be used to identify organisms (as discussed above, biomaterials or biological samples comprising nucleic acids are obtained from the organism) ). The teachings are also directed to kits for extracting nucleic acids from a matrix, including but not limited to, calcified or hardened biological materials, materials believed to contain adhesion or adhesion of nucleic acids, and denim considered to contain nucleic acids. . Such kits can be used for archaeological, criminal, forensic, human identification 099131421 疋 and research using the methods described above. In some embodiments, the basic kit can comprise a 4-, adhesive and/or viscous material or denim material having a cleavage/excitation capable of releasing nucleic acids from the matrix. The kit may also optionally include (4) instructions. This kit may also contain other optional kit components, such as, for example, Form No. A0101 帛 34 pages / Total 54 pages containing polymer and cleaning solutions for 1003014903-0 201213805, magnetic suction particles, or More cleaning solutions, nuclear loss extraction solutions, nucleic acid extraction control groups, magnetic devices, and educational programs and manuals that limit misuse. The number of sub-types and magnetic devices in the set can also vary depending on several factors (e.g., the sensitivity of the most official/process). Within the scope of these teachings, test kits for manual testing or test kits for use in the preparation, use, detector or analyzer are used. Those in the field understand that the technology used is generally not limited (4)

. Conversely, a variety of nucleic acid extraction methods are within the "methods and sets" of the disclosed combination (assuming a sub-(four) bribe or a nucleic acid with a material). ;

Although the principles of the present invention have been described in connection with the specific embodiments of the present invention, it should be understood that the description is not intended to limit the scope of the invention. It has been disclosed herein for the purposes of illustration and description. It is not intended to be exhaustive or to limit the invention to the precise form. Many modifications and variations will be apparent to those skilled in the art. The selection and description of the present invention are intended to best explain the principles and practice of the specific embodiments disclosed herein. The scope of the present disclosure is to be understood as being limited by the following examples, which are not to be construed as limiting the scope of the present teachings. Those skilled in the art will recognize that many modifications, substitutions, and equivalents are possible. All such modifications, substitutions, and equivalents are 099131421 Form No. A0101 Page 35 of 54 1003014903-0 201213805 Thoughts are included herein. The following steps of the acid counteract the n- <agent and the steps that can be used to extract nucleic acids from the core substrate and the susceptor containing the nuclear 15 biomolecule. The sample pretreatment uses mild Clean, work s Valley liquid air-dried overnight, and use the connection to Dremel, the grinding stone to remove the bone and teeth ', layer (D Remel, Madison, WI) and clear, ° & large pieces of bone are cut into small pieces. Use fresh or moon-cleaned grinders (eg pr〇da Cryo-mill, BioSpect ucts, Bart 1·, 磨Recognize x esviUe, OK) Grind the bone and the tooth block. Store the grinding, month or tooth powder in the cigarette butt that will be inserted into the cigarette butt, and (the room temperature; remove the filter material and contact the skin V as a finger The first 5 mm part of the bottom of the private paper of J is cut into three. The sample of the scented gum is flattened and stored at room temperature. The glue is stored in a clean culture dish: 3 (four) thick In order to harden, the lid is closed and will be cultured | eight; 0 C for several hours. The hard-top candied into 8 parts of the same size is used as the observation of the extraction. Samples of peeling and peeling from various sources are prepared. Use 1.5 per sample

Cm Υ 〇 r 2.5 cm block tape (including biological samples). The blood or saliva is dried and then added to the lysis reagent solution prior to removal by stripping. For tape + biological fluid samples, a night of 5_5 ^ or saliva is added directly to the side of the adhesive on the tape. For tape stripping samples, add 2 human blood or 5 A saliva to the slide. When the biomaterial is dry, the biomaterial is stripped with tape. A tape peeling sample containing skin cells is also collected from either the inside of the shirt collar of the passed sputum sample or the inside of the shirt cuff. 099131421 Form No. A0101 Page 36 of 54 1003014903-0 Prepare a sample of twill fabric (non-black denim). Blood was replicated 4 times using a 5 mm perforation where the blood was broken and air dried. ★ The lysis reaction with the Tris-C Bubuffer lysis reagent solution was applied to the LySepTM tube (Applied Biosystems) in a volume of 5 〇〇uL. The & column was incubated in a temperature mixer at 56 C for 5 minutes in the presence of 3 L reducing agent (1 μ) and 3 uL 2 0 哗 / this enzyme. The DMA extraction was done using the AutoMate ExpressTM kit (Applied iosystems) according to the manufacturer's plan. Use the Quantifiler DuoTM Set (Applied Bi〇systems) to complete the DNA quantification according to the manufacturer's instructions. Use the Jderitifiler kit to perform the STR profile according to the manufacturer's instructions and follow Applicator's instructions to AppHed.

The CE analysis was performed on the Bl〇systems 3130x1. As described for the blood on the denim, a coarse twill fabric sample prepared by analyzing the oral epithelial cells of 6 copies of the oral cavity was analyzed. The blood on the black denim was 2 uL of 3 replicates of blood and was analyzed as described above for the denim. Preparation of ΐι·Cleavage Reagent Solution A sufficient amount of lysate solution is prepared to treat the number of samples of secondary or tertiary replication. Via 220 liters of 250-500 sewed chelating agent, at least one detergent containing U w / v (added to 〇 5_ι mg / mL of enzyme)? 117.9 to 8.5 and the final concentration is 1〇_1〇〇龙. The freshly prepared reducing agent is mixed in DNase water and freshly prepared. The lysate mixture was incubated at a temperature range of 450-70 C plus or without shaking for a period of 9 minutes to 2 hours. Table 6 illustrates the bribery solution that can be used to extract from solid biomaterials and return-to-financial materials. The solid biofilm 4 and the return material contain or be acknowledged with a nuclear-containing form number. Page 1 of 54 Page 100 of 1003 (201213805 Acid biological sample. 111. Preparation of Tris-n-buffered lysis reagent solution Prepare a sufficient amount of Tris-n-buffered lysing reagent solution to treat the number of samples of secondary replication, triple replication or 6 replications. 220 μί of 250-1 000 mM chelating agent, containing 0.5 - 3% w/v of at least one detergent (to which 0.5-50 mg/mL of enzyme is added), pH 7. 9 to 8.5 and the final concentration is 10-1 00 mM of the freshly prepared reducing agent in DNase-free water is mixed and the solution is freshly prepared. The cleavage mixture is incubated at a temperature range of about 50-70 ° C for a period of 90 minutes to 2 hours with or without shaking. Table 6: Split Pit Formula 1 Formulation 配方 Formula HI Formulation IV Formulation V Formulation W Formulation vn EGTA 250 to 500 mM 0 250 to 500 mM 0 250 to 500 mM 250 to 500 mM 250 to 500 mM EDTA 0 250 to 500 mM 0 250 to 500 mM 0 0 0 DTT 10 to 100 mM 10 to 100 mM 10 to 100 mM 10 to 100 mM 10 to 100 mM 10 to 100 mM 10 to 100 mM NLS 0.5 to 3% w/v 0.5 to 3 % w/v 0 0 0.5 to 3 % w/v 0 0 deoxycholic acid Pin 0 0 0.5 to 3 % w/v 0·5 to 3 % w/v 0.5 to 3 % w/v 0 0.5 to 3 % w/v SDS 0 0 0 0 0 0.1 to 0.5% «^ 0 0 0 0 0 0 50 to 150 mM 0 Triton X 100 0 0 0 0 0 0.1 to 0.3 % v/v 0 NP-40 0 0 0 0 0 0 0-1 to 0.3% Protease κ 0.5 to 1 mg/mL 0.5 To 1 mg/mL 0.5 Μ 1 mg/mL 0.5 Μ 1 mg/mL 0.5 to 1 mg/mL 0.5 Μ 1 mg/mL 0.5 to 1 Table 7: Washing solution: Group composition 10 mM Tris-CI, pH 8.0 0.1 mM EDTA 95% sodium alcohol deoxycholate (DSD)

Cleaning buffer A

70% 0.5% Clear B earns 4 mM 0.04 mM 60% Final volume

100 mL 100 mL III. Extraction of nucleic acids from solid biological samples A freshly prepared lysis solution of 230 was added to 50-200 mg of bone or 10 mg of tooth powder in a 2.0 mL screw cap. Close the lid 099131421 Form No. A0101 No. 38 Buy / Total 54 Pages 1003014903-0 201213805 Closed and placed at 56 C in Eppendorf® Thermomixer R (

Incubate at 1100 rpm for 2 hours in Eppendorf North America, Westbury, NY. At the end of the incubation, the tube was centrifuged at 1 〇, 〇〇〇X g for 5 minutes' and the clarified supernatant/lysate (up to 21 () L·) was transferred to a lysis buffer containing 3 〇〇pL prepFilerTM In a new 1.5 mL centrifuge tube, the DNA was separated according to the manufacturer's large volume book. Basically, 15 uL of PrepFilerTM magnetic particles were added with uL 99.5% isopropanol, followed by gentle mixing. This pellet was washed twice with PrepFilerTM Wash Buffer (which used 6 μL of L and then 300 μL of buffer a, followed by 30 μL of PreLerTM Wash Buffer B). Following the book and the PrepFiler Automated Forensic DNA Extraction Kit or the prepFUer Manual Kit User Guide (P/N 4390932-01), the nucleic acid was flushed to 5 〇 UL's PrepFilerTM Extraction Buffer. IV. Extraction of nucleic acids from adhesive materials

A 23 〇 wL freshly prepared lysis solution was added to the sample containing adhesive prepared in the 5.5 ‘centrifuge tube as described above. The lid was closed and the tube was incubated at 56 ° C for 4 minutes at 750 rpm in an Eppendorf® Thermometer. At the end of the incubation, the contents were transferred to a filter column (Axygen Scientific, Inc., Union City, CA). The supernatant/lysate was separated from the substrate by centrifuging the filter column at i6, 110 x g for 2 minutes. 300 pL of PrepFiler "* lysis buffer was added to the clarified supernatant/lysate and DNA isolation was performed according to the manufacturer's protocol. V. Extraction of nucleic acids from denim material A freshly prepared lysis solution of 500 is added to a sample containing the sample prepared in the 1.5 mL LySep 099131421 Form No. A0101 Page 39/54 Page 1003014903-0 201213805 tube as described above. Sex (fluid). The lid was closed and the tube was incubated at 56 ° C for 40 minutes at 750 rpm in an Eppendorf® Thermo-mixer R. The supernatant/lysate was separated from the substrate by centrifugation for 2 minutes at ,16, 110 X g. 300 UL PrepFilerTM Lysis Buffer was added to the clarified supernatant/lysate and manufactured according to The program is DNA-separated. VI. DNA isolation is as described by the manufacturer (Applied Bi〇systems, Prep_ FilerTM Forensic DNA Extraction Kit User Guide. pn 4390932 Rev. B, (2008)) DNA from lysates was isolated from a large sample of studies (for the use of 500 PrepFilerTM Lysis Buffer). VII. Human DNA quantification is as described in the user manual (Applied Biosystems, Quanti fi ler® Human DNA Quantitation Kit and Quantif丨ier® Y Male Human DNA Quantitative Set.: :: User's Manual _.: vpp:N 4344790

Rev. B, (2003)), using a DNA extract of 2 WL, the amount of DNA was determined via a Quant i filer icQuanti filer DUO® human DNA quantification kit (Applied Biosystems, Foster City, CA). PCR was performed on a 7500 real-time PCR system and analyzed using 7500 System SDS software vl.2.3 (Applied Biosystems, Foster Ci ty, CA). VIII. Short tandem repeat (STR) analysis using the steps described in the User Manual (Applied Biosystems, AmpF STR® Identi Fi ler® PCR Amplification Kit. User Manual. ppN 4323291 Rev. B, (2001)), The DNA extract obtained from the biological sample will be amplified for use in the Identi filer suite analysis using the PrepF iler 099131421 Form No. A0101 page 40/54 page 1003014903-0 201213805TM forensic DNA extraction kit. As in the User Guide (Applied Biosystems, ABI PRISM 2 3100 Genetic Analyzer and ABI PRISM® 3100 - Avant Genetic Analyzer User Reference Guide, PN 4335393. RevA, (2002) 'Appl ied Biosy stems for Human Identification Applications The products described in GeneMapper® ID Software) were analyzed using the GeneMapper® ID Software v3. 2.1 and analyzed at 3100 or 313Ox 1 Genetic Analyzer (Applied Biosystems). While the present disclosure has been described with respect to various specific embodiments and examples, various modifications may be made without departing from the spirit and scope of the invention. BRIEF DESCRIPTION OF THE DRAWINGS [0006] Fig. 1 is a diagram depicting the inter-color balance (Intercolor balance (ICB)) between STR dual genes for repeated blood extracted from denim. The ICB is based on each color. The maximum peak is divided by the minimum peak and counted as 0 (the peak of the heterozygote is the average and the homozygote is divided by 2). The samples are listed in Table 6. [Main component symbol description] [0007] ICB color balance 099131421 Form number A0101 Page 41 of 54 1003014903-0

Claims (1)

201213805 VII. Patent application scope: ^ A composition for solving a solid tissue, the composition comprising a detergent, a chelating agent, a reducing agent and one or more of an enzyme. 2. The composition of claim 1, wherein the cleaning agent is selected from one or more of the following: N_lauric myosin, sodium dehydrocholate jTAB, dodecyl hydrazine _D-maltose, thiol-methyl branamide, polyethylene glycol P-(l,l,3,3-tetradecylbutyl)-phenyl ether, sodium lauryl sulfate and Conjugate. 3. The composition of claim 1, wherein the chelating agent is one or more of the following: ethylene glycol tetraacetate (EGTA), ethylenediaminetetraacetate (EDTA), and The citric acid and its combination. 4. If you apply for a patent scope! The composition according to the above, wherein the reducing agent comprises one or more of the following: tris(2-sulfate phosphine (TCEP), sulfur red alcohol (DTE), and dithiothreitol ( DTT) 5. The composition of claim W, wherein the enzyme is T or less than T: caspase, chymotrypsin, lunar protease, protease ash , thrombin, staphylococcus, protease, protease, papain, bacillus, bacterium, bacterium, bacterium, bacterium, bacterium, bacterium, bacterium, bacterium, bacterium, bacterium, bacterium, bacterium, bacterium, bacterium, bacterium, bacterium, bacterium, bacterium The composition of claim 6, wherein the solid tissue is a calcified tissue, a biomaterial, an adhesive material or an adhesive substrate. 8. The composition of claim 7 The composition of the biological material is selected from the group consisting of bone, cartilage, ligament, tendon, and tooth. 099131421 Form No. A0101 Page 42 of 54 1003014903-0 201213805 y. The composition of claim 7, wherein The adhesive material comprises The composition of claim 9, wherein the biological sample is attached to the adhesive material. The composition of claim 7, wherein the composition of claim 7 The adhesive material is selected from the group consisting of chewing gum, cigar butt, cigarette butts, adhesive films, adhesive labels, stickers, skin patches, envelopes, envelope seals, stamps, fingerprint tape peeling and tape, duct tape and electrical tape. 12. The composition of claim 11, wherein the dermal patch is selected from the group consisting of: an electronic electrode, a transferable totem, a skin infiltrated chemical patch, a wound A method of making a product, wherein the product comprises a nucleic acid, the method comprising: a) culturing a solid in a solution; b) extracting a supernatant, wherein The supernatant contains the nucleic acid extracted from the solid. The method of claim 13, wherein the solid comprises the nucleic acid. The method of claim 14, wherein the solid is a raw material or an adhesive material or a substrate. The method of claim 15, wherein the biological material is selected from the group consisting of a reduced tissue, a bone, a cartilage, a ligament, a tendon, and a tooth. The method of claim 15, wherein the adhesive material comprises a biological sample. 18. The method of claim 17, wherein the biological sample 099131421 Form No. A0101 Page 43 of 54 1003014903-0 201213805 is incorporated into the adhesive material. 19. The method of claim 18, wherein the adhesive material is removed from the git' (10) buttocks, the adhesive film, the adhesive label, the skin patch, the stamp, and the fingerprint tape. The method of claim 19, wherein the skin patch is selected from the group consisting of: an electronic electrode, a transferable totem, and a skin infiltrated chemical patch. Injury π care dressing. The method of claim 13, wherein the solution comprises one or more of the following: a detergent, an integrator, a reducing agent, and an enzyme. The method of claim 21, wherein the cleaning agent is selected from the group consisting of 卩 or more: 卩 _ laurel muscle, sodium deoxycholate, CTAB, twelfth stone _D _ maltose, thiol_Ν_methyl branamide, polyethylene glycol oxime-(1,1,3,3-tetramethylbutyl)-phenyl ether, sodium lauryl sulfate and Conjugate. The method of claim 21, wherein the chelating agent comprises at least one of the following: tetraacetic alcohol (EGTA), ethylenediamine tetraacetate (EDTA), and citric acid. The method of claim 21, wherein the reducing agent is at least one of the following: tris(2_cycloethyl)phosphine (TCEp), dithioerythritol (DTE) And di-threitol (j) TT). 099131421. The method of claim 21, wherein the enzyme comprises one or more of the following: caspase, chymotrypsin, pepsin, protease, thrombin, Staphylococcus V8 protease, pronase, papain, Bacillus sp. E1A protease and trypsin are listed in the form No. A0101 1003014903-0, page 44/54, 201213805 and combinations thereof. A method of producing a product, wherein the product is a nucleic acid, the method comprising the steps of: (a) culturing the same solution as a solution; (b) shaking the culture solution; (c) extracting A supernatant product, wherein the nucleic acid is located in the supernatant. The method of claim 26, wherein the sample comprises the nucleic acid. 〇 28. The method of claim 27, wherein the sample is a biomaterial, an adhesive material or a substrate, or a natural or synthetic substrate. The method of claim 28, wherein the biological sample is selected from the group consisting of bone, cartilage, ligament, tendon, and tooth. 3. The method of claim 28, wherein the adhesive material comprises a biological sample. The method of claim 30, wherein the biological sample is attached to the adhesive material. The method of claim 31, wherein the adhesive material is selected from the group consisting of chewing gum, cigar butt, cigarette butt, adhesive film, adhesive label, sticker, skin patch, envelope, stamp, fingerprint tape The method of claim 32, wherein the skin patch is selected from the group consisting of an electronic electrode, a transferable totem, and a skin infiltrated chemical patch. And wound care dressings. The method of claim 26, wherein the solution comprises one or more of the following: a cleaning agent, a chelating agent, a reducing agent 099131421, Form No. A0101, page 45/54 Page 1003014903-0 201213805 and an enzyme. The method of claim 34, wherein the detergent is selected from one or more of the following: N-lauric acid, sodium deoxycholate, CTAB, dodecaine -D-maltose, thiol-N-methyl branamide, polyethylene glycol p-(l,1,3,3-tetramethylbutyl)-phenyl ether, sodium lauryl sulfate And its combination. The method of claim 34, wherein the chelating agent comprises at least one of the following: ethylene glycol tetraacetate (EGTA), ethylenediamine tetraacetate (EDTA), and citric acid. 37. The method of claim 34, wherein the reducing agent comprises at least one of tris(2-carboxyethyl)phosphine (TCEP), dithioerythritol (DTE), and Dithiothreitol (DTT). 38. A method of making a product, wherein the product comprises a nucleic acid, the method comprising: (a) lysing a solid sample in a lysis solution to produce a lysate comprising one of the nucleic acids; (b) extracting The cleavage product, wherein the nucleic acid system is located in the lysate, the method of claim 38, wherein the solid sample comprises the nucleic acid. 40. The method of claim 39, wherein the solid is a biomaterial, an adhesive material or a substrate, or a natural or synthetic substrate. The method of claim 40, wherein the biological material is selected from the group consisting of bone, cartilage, ligament, tendon, and tooth. 42. The method of claim 40, wherein the adhesive material substrate comprises a biological sample. 099131421 Form No. A0101 Page 46 of 54 1003014903-0 201213805 43. The method of claim 42, wherein the biological sample is embedded in the adhesive material substrate. 44. The method of claim 43, wherein the adhesive material is selected from the group consisting of chewing gum, cigar butt, cigarette butts, films, adhesive labels, stickers, skin patches, envelopes, stamps, and tapes. The method of claim 44, wherein the skin patch is selected from the group consisting of an electronic electrode, a transferable totem, a skin infiltrated chemical patch, and a wound care dressing. . 46. The method of claim 38, wherein the lysis solution system comprises one or more of the following: a cleaning agent, a chelating agent, a reducing agent, and an enzyme. 47. The method of claim 46, wherein the detergent is selected from one or more of the following: N-lauric myosin, sodium deoxycholate, CTAB, twelve base points - D-maltose, thiol-N-mercapto branamine, polyethylene glycol p-(l, 1,3,3-tetramethylbutyl)-phenyl ether, sodium lauryl sulfate and Its combination. The method of claim 46, wherein the chelating agent comprises at least one of the following: ethylene glycol tetraacetate (EGTA), ethylenediaminetetraacetate (EDTA), and citric acid. . The method of claim 46, wherein the reducing agent comprises at least one of tris(2-carboxyethyl)phosphine (TCEP), dithioerythritol (DTE), and Dithiothreitol (DTT). 50. The method of claim 46, wherein the enzyme comprises one or more of the following: caspase, chymotrypsin, pepsin, proteinase K, thrombin, grapes Cocinobacterium V8 protease, pronase, papain, Bacillus sp. El A protease and TGase are 099131421 Form No. A0101 Page 47 / Total 54 Page 1003014903-0 201213805 and combinations thereof. 51. A method of isolating a nucleic acid from a solid sample, the method comprising: (a) lysing the solid sample in a lysis solution to produce a mixture comprising the nucleic acid; (b) the mixture Centrifugation wherein the nucleic acid is separated from the solid sample. The method of claim 51, wherein the solid sample comprises the nucleic acid. 53. The method of claim 52, wherein the solid is a biomaterial, an adhesive material or a substrate, or a natural or artificial substrate. 54. The method of claim 53, wherein the biological material is selected from the group consisting of bone, cartilage, ligament, tendon, and teeth. The method of claim 53, wherein the adhesive material comprises a biological sample. The method of claim 55, wherein the biological sample is attached to the adhesive material. 57. The method of claim 56, wherein the adhesive material is selected from the group consisting of chewing gum, cigar butt, cigarette butt, adhesive film, adhesive label, sticker, skin patch, envelope, stamp, and fingerprint tape stripping. The method of claim 57, wherein the skin patch is selected from the group consisting of an electronic electrode, a transferable totem, a skin infiltrated chemical patch, and Wound care dressing. The method of claim 51, wherein the lysing solution comprises one or more of the following: a cleaning agent, a chelating agent, a reducing agent, and an enzyme. The method of claim 59, wherein the cleaning agent is selected from one or more of the following: N-lauric muscles, the method of claim 59, wherein Amino acid, sodium deoxycholate, CTAB, dodecyl yS-D-maltose, thiol-N-methyl branamide, polyethylene glycol p-(l,1,3,3-tetra Butyl)-phenyl ether, sodium lauryl sulfate and combinations thereof. 61. The method of claim 59, wherein the chelating agent comprises at least one of the following: ethylene glycol tetraacetate (EGTA), ethylenediamine tetraacetate (EDTA), and citric acid. 62. The method of claim 59, wherein the reducing agent comprises at least one of: tris(2-carboxyethyl)phosphine (TCEP), dithioerythritol (DTE) And dithiothreitol (DTT). 63. The method of claim 59, wherein the enzyme comprises one or more of the following: caspase, chymotrypsin, pepsin, proteinase K, thrombin, grapes Coccus V8 protease, pronase, papain, Bacillus sp. sp. E1A protease and TGase and combinations thereof. ^ 64 . A nucleic acid product produced by the method described in claim 13 of the patent application. Q 65. A nucleic acid product produced by the method described in claim 26 of the patent application. 66. A nucleic acid product produced by the method described in claim 38. 67. A nucleic acid product isolated by the method described in claim 51. 68. A method of identification comprising: (a) lysing a cleavage solution in a lysis solution to produce a lysate comprising nucleic acids from one of said samples; (b) extracting said lysate, wherein The nucleic acid is located in the lysate 9 (C) to analyze the nucleic acid; 099131421 Form No. A0101 Page 49 / Total 54 Page 1003014903-0 201213805 69 70 71 72 73 74 75 76 099131421 (d) Identification based on nucleic acid analysis The sample. The fingerprint tape was peeled off according to the method described in claim 76. The method of claim 51, wherein the lysis solution 包含 comprises one or more of the following: a coffee agent and an enzyme. The method of claim 59, wherein the cleaning agent is selected from the group consisting of - or more: N_lauric acid, sodium deoxycholate, CTAB, twelfth terminal maltose Gan, thiol. methyl branamide, polyethylene glycol P-(1,1,3,3-tetramethylbutyl)-benzoic acid, sodium lauryl sulfate and combinations thereof. The method of claim 59, wherein the chelating agent is at least four of ethylene glycol (E.g.), ethylenediaminetetraacetate (EDTA) and citric acid below the package 3. The method of claim 59, wherein the reducing agent comprises at least one of the following: a tris(2-aged) scale (TCEp), a dithioerythritol (DTE), and a disulfide. Glycocalyx (four) hit). The method of claim 59, wherein the enzyme comprises one or more of the following: caspase, chymotrypsin pepsin, protease kappa, thrombin, Staphylococcus V8 Protease, pronase, papain, Bacillus sp. EU protease and trypsin and combinations thereof. The method of claim 69, wherein the step of comparing comprises directing the fingerprint with a fingerprint collection library. A kit for extracting nucleic acids from a solid sample, the kit comprising: a lysis buffer. Form No. Α0101, wherein the sample is a chelating agent, also 50th page/54 pages 10030l·201213805 77.78.79. Ο80.81. Ο82.83.84. Set according to claim 76 The group, wherein the lysis buffer comprises one or more of the following: a detergent, a chelating agent, a reducing agent, and an enzyme. The kit of claim 76, wherein the cleaning agent is one or more selected from the group consisting of strontium-lauric acid, sodium deoxycholate, CTAB, and twelfth/SD. - maltose, thiol-oxime-methyl branamide, polyethylene glycol ρ-(1,1,3,3-tetramethylbutyl)-phenyl ether, sodium lauryl sulfate and Conjugate. The kit of claim 76, wherein the chelating agent comprises at least one of the following: ethylene glycol tetraacetate (EGTA), ethylenediamine tetraacetate (EDTA), and citric acid. The kit of claim 76, wherein the reducing agent comprises at least one of: tris(2-carboxyethyl)phosphine (TCEP), dithioerythritol (DTE), and Thiothelitol (DTT). The kit of claim 76, wherein the enzyme comprises one or more of the following: caspase, chymotrypsin, pepsin, proteinase K, thrombin, staphylococcus It belongs to V8 protease, pronase, papain, Bacillus sp._E__1__A protease and trypsin and combinations thereof. The kit of claim 76, wherein the solid sample is selected from the group consisting of a biomaterial and an adhesive material. The kit of claim 82, wherein the biomaterial is selected from the group consisting of bone, cartilage, ligament, tendon, and teeth. The kit of claim 83, wherein the adhesive material is selected from the group consisting of chewing gum, cigar butt, cigarette butt, adhesive film, adhesive label, sticker, skin patch, envelope, stamp, fingerprint tape stripping, and the like. Tape 099131421 Form No. A0101 Page 51 of 54 1003014903-0 201213805 85. The kit of claim 84, wherein the skin patch is selected from the group consisting of: an electronic electrode , transferable totems, skin penetration chemical patches, and wound care dressings. 86. The kit of claim 76, which optionally comprises: a solution comprising a polymer and a cleaning agent; and particles that are magnetically attracted. 87. The kit of claim 86, wherein the polymer comprises one or more of the following: dextran, cellulose, cellulose derivatives, soluble starch, dextrin, fiber Dextrin, polyethylene glycol, heparin, glycogen, and combinations thereof. 88. The kit of claim 86, wherein the detergent comprises one or more of the following: N-lauric myosin, sodium deoxycholate, CTAB, dodecaine -D-maltose, thiol-N-mercapto branamine, polyethylene glycol p-(l,1,3,3-tetramethylbutyl)-phenyl ether, sodium lauryl sulfate And its combination. 89. The kit of claim 86, wherein the magnetically permeable particles comprise magnetic nanoparticles coated with dextran. 90. The kit of claim 86, wherein all of the contents of the on-demand solution are in a single container. 91. The kit of claim 86, further comprising a magnetic device. 92. The kit of claim 86, further comprising a cleaning solution. 93. The composition of claim 1, further comprising a salt selected from the group consisting of KCl's NaCl, MgCl, and MnCl2. 94. A composition for lysing a biological sample on a denim, the 099131421 Form No. A0101 Page 52 of 54 1003014903-0 201213805 The composition comprises one or more of the following: a cleaning agent , a chelating agent, a salt, a reducing agent and an enzyme. The composition of claim 94, wherein the detergent is selected from one or more of the following: N-lauric myosin, sodium deoxycholate, CTAB, ten Di-male maltose, thiol-N-methyl branamide, polyethylene glycol p-(l,1,3,3-tetramethylbutyl)-phenyl ether sodium lauryl sulfate and Its combination. 96. The composition of claim 94, wherein the chelating agent comprises one or more of the following: ethylene glycol tetraacetate (EGTA), ethylenediamine tetraacetate (EDTA) ) with citric acid and its combination. The composition of claim 94, wherein the reducing agent comprises one or more of the following: tris(2-carboxyethyl)phosphine (TCEP), dithioerythritol (DTE) And dithiothreitol (DTT). 98. The composition of claim 94, wherein the enzyme comprises one or more of the following: caspase, chymotrypsin, pepsin, proteinase K, thrombin, Staphylococcus genus 8 protease, pronase, papain, bacillus sp. E1A protease and trypsin 〇 99. The composition of claim 94, wherein the denim comprises the nucleic acid. 100. A sleeve containing a lysis buffer as described in claim 94 of the patent application. 099131421 Form No. A0101 Page 53 of 54 1003014903-0