CN102229927A - Method and reagent for improving rate of extraction of DNAs of castoff cells in case trace sample - Google Patents
Method and reagent for improving rate of extraction of DNAs of castoff cells in case trace sample Download PDFInfo
- Publication number
- CN102229927A CN102229927A CN2011101488697A CN201110148869A CN102229927A CN 102229927 A CN102229927 A CN 102229927A CN 2011101488697 A CN2011101488697 A CN 2011101488697A CN 201110148869 A CN201110148869 A CN 201110148869A CN 102229927 A CN102229927 A CN 102229927A
- Authority
- CN
- China
- Prior art keywords
- dna
- liquid
- conjunction
- magnetic bead
- peg
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Images
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention relates to a method and a reagent for improving the rate of extraction of the DNAs of castoff cells in a case trace sample. Due to the problems of trace amount of case sample, keratinization of cells, multiple polymerase chain reaction (PCR) complex amplification of 16 short tandem repeat (STR) loci and the like, the extraction of the DNAs of castoff cells has long been a difficult point in forensic medicine. According to researches, the combination of pronase digestion, bath at 70 to 100 DEG C and polyethylene glycol(PEG)-ethanol magnetic bead combined system can help to effectively extract trace DNAs and to obtain DNA fragments with a proper length, so that the need of complex amplification of the 16 STR loci in case detection can be satisfied and the success rate of the case detection is improved. The pronase digestion and warm bath allow the castoff cells to release DNAs effectively and ensures the relatively complete structure of the DNAs and the easy combination of the DNAs with the magnetic beads in the PEG-ethanol system. The use of the PEG-ethanol combined system promotes the high-efficiency DNA adsorption of the magnetic beads, ensures the relatively complete structure of the DNA and ensures that DNA fragments with proper length can be obtained to satisfy the need for the complex amplification of the 16 STR loci in case detection.
Description
Technical field
The present invention relates to the method and the reagent of case trace sample cast-off cells DNA high efficiency extraction, belong to forensic DNA detection field.
Background technology
Since round pcr is applied to the check of biological material evidence, after especially using the str locus seat and analyzing, DNA extraction just becomes the key link of forensic dna polymorphism analysis, and the quality of DNA sample quality will be directly connected to the success or failure of subsequent experimental.But in actual inspection case, increasing case is because the effect of the character of case own, commit a crime course and offender's factors such as counterreconnaissance consciousness enhancing, in some site inspections, be difficult to find conventional biological materials such as blood cake, seminal stain, hair, cigarette end, the sample that therefore may be loaded with human body case trace sample just becomes the important material evidence of cracking of cases.
Human body case trace sample amount is little, and mostly is keratinization, and how nucleus degenerates, on the various carriers that contact with human body.Its carrier has following characteristics: 1. sample big area such as micro-epidermic cell is distributed in carrier surface and combines closely with carrier, dirt, is difficult to collect shift; 2. the shelf time long or improper, DNA easily degraded is difficult to satisfy 16 STR site multiplex PCR composite amplifications (16 templates increase simultaneously in the pipe); 3. this type of sample carrier is many through washing, contains the PCR inhibition.Therefore, the extraction difficulty of case trace sample DNA is big, lacks stable extracting method, causes the failure of inspection case easily.
At present, according to the amount of case trace sample and the difference of its place carrier, the method for DNA extraction is also different.The more sample of case trace sample that people's contact levels such as clothing, footwear, socks are big, may contain, behind many case trace samples by case trace sample absorption apparatus collection sample, extract with automatically working station or chelex-100 method, can be by methods such as prolongation incubation period, the amount that increases Proteinase K, minimizing elution volumes, to reach the purpose that improves dna profiling concentration.And when the wooden handle that runs into various tool, as the wooden handle of axe, the fingerprint on all kinds of carrier,, carrier less as the human body contacts site such as finger mark on the bowl cover is during to the stronger sample of case trace sample adsorptivity, the reviewer takes the method for different extractions according to the extraction experience of oneself, but often all is that the intersection use of plurality of reagents box and the mixing of multiple extracting method are used.As hatching several hrs with Chelex-100 and Proteinase K after Microcon-100 purification column purifying concentration of DNA; Or extract dna profiling with other test kits such as QIAGEN M48 magnetic bead kit, have also wherein with chloroform purification step utilization in organic method.The DNA amount that these methods are extracted less, purity is low, length is imperfect, not only will use expensive import reagent, and complex steps, length consuming time, complicated operation, personal experience influence greatlyyer, and success ratio is lower.
The inventor optimizes the method and the reagent of cracking, articulated system invention raising case trace sample DNA extraction efficient, solves the above problem that runs in the DNA extraction process, and is successfully applied to actual inspection case.
Summary of the invention
The technical problem to be solved in the present invention is, system optimization cracking, combination, cleaning system solve an extraction difficult problem that only contains minim DNA in the case sample, realizes easyly, quick, extracts minim DNA efficiently.
Through different condition combination, serial simultaneous test and a large amount of on-the-spot case sample test, we find: 1. protease digestion, the bath of 70-100 temperature, PEG-ethanol magnetic bead can the high efficiency extraction minim DNAs in conjunction with the combination of 3 of liquid; 2. protease digestion, temperature are bathed to handle and can efficiently be discharged cast-off cells DNA, and structural integrity easily combines with magnetic bead in the PEG-ethanol system; 3. the use of PEG-ethanol articulated system not only promotes the efficient absorption of magnetic bead to DNA, and has guaranteed that dna structure is complete relatively, the dna fragmentation that obtains to be of convenient length can satisfy case detect in the needs of 16 STR site composite amplifications.Reagent extracts case trace sample DNA can either reduce cost, easy operation, and the intersection that need not plurality of reagents box and several different methods is used, and highly sensitive, and method is simple, has reduced the empirical dependence to the technician, has realized that case is detected as the lifting of power.
The invention provides a kind of reagent that extracts case trace sample DNA, it consists of the magnetic bead suspension; Lysate, it consists of the Tris-HCl damping fluid of pH=8.0; In conjunction with liquid I, it consists of the PEG of 20-50%w/v, the MW=8000 of PEG; In conjunction with liquid II, it consists of ethanol; Scavenging solution, it consists of the ethanolic soln of 50-80%v/v.
Of the present invention can be with a kind of replacement ethanol in methyl alcohol and propyl alcohol, the Virahol in conjunction with liquid II.
The inventive method can be extracted DNA from micro-case sample in simple and effective ground.
Micro-case sample of the present invention is the biological material that contains micro-case trace sample that criminal-scene is left over.
The carrier that has shifted case trace sample of the present invention is resulting filter membranes such as the clothing that adsorbs criminal-scene with the biomass cells extraction apparatus and leave over, rope, electric wire, waddy.
The carrier that has shifted case trace sample of the present invention is resulting glued membrane or scotch tapes such as fingerprint on clothing, rope, electric wire, waddy, bottleneck, various handle or handle, glass or the desktop etc. left over case trace sample Adhesive-fetch device, the sticking enchashment field of scotch tape.
The carrier that has shifted case trace sample of the present invention is resulting filter paper, cotton swab, the yarns etc. such as fingerprint on the sassafras scenes of wiping away such as filter paper, cotton swab, yarn clothing, rope, electric wire, waddy, bottleneck, various handle or handle, glass or the desktop etc. left over.
Be used for lysate of the present invention, in conjunction with liquid I, be not particularly limited in conjunction with liquid II, scavenging solution, the lysate that can adopt for this area, in conjunction with liquid I, in conjunction with liquid II, scavenging solution.
Major advantage of the present invention is:
1. the conjugated protein enzymic digestion of the inventive method, the bath of 70-100 degree temperature can fully cracking case trace sample cast-off cells, and the dna structure of release is complete relatively, in the PEG-ethanol system, easily combine with magnetic bead and;
2. PEG-ethanol not only can be removed protein better in conjunction with the existence of liquid during reagent was formed, and during magnetic bead can be well-dispersed in conjunction with liquid when adsorption of DNA, had realized that magnetic bead is to the efficient absorption in conjunction with DNA in the liquid;
3.PEG-ethanol is fit to the DNA length of extracting in conjunction with the use of liquid, can satisfy the needs of 16 STR site composite amplifications in the case detection, greatly reduces and loses peak, amplification energy imbalance.
4. this extracting method has reduced the empirical dependency to the operator, and is easy and simple to handle, good stability;
5. need not to use the big organic solvents of toxicity such as phenol, chloroform, security is good;
6. highly sensitive, to extract fast, general operation can be finished at 1～1.5 hour, and can be used for the DNA extraction at automatically working station.
Description of drawings
The composite S TR amplification collection of illustrative plates of the minim DNA that the belt that Fig. 1 criminal-scene is left over extracts.
The DNA composite S TR amplification collection of illustrative plates that Figure 21 μ L extracts through 60 times of dilution fresh bloods.
Figure 31 μ L is through 60 times of dilution fresh bloods, and DNA composite S TR amplification collection of illustrative plates is extracted in control experiment 2.
Figure 41 μ L is through 60 times of dilution fresh bloods, and DNA composite S TR amplification collection of illustrative plates is extracted in control experiment 3.
Figure 51 μ L is through 60 times of dilution fresh bloods, and DNA composite S TR amplification collection of illustrative plates is extracted in control experiment 4.
Figure 61 μ L is through 60 times of dilution fresh bloods, and DNA composite S TR amplification collection of illustrative plates is extracted in control experiment 5.
Figure 71 μ L is through 60 times of dilution fresh bloods, and DNA composite S TR amplification collection of illustrative plates is extracted in control experiment 6.
Figure 81 μ L is through 60 times of dilution fresh bloods, and DNA composite S TR amplification collection of illustrative plates is extracted in control experiment 7.
Figure 91 μ L is through 60 times of dilution fresh bloods, and DNA composite S TR amplification collection of illustrative plates is extracted in control experiment 8.
The slippers that Figure 10 leaves over for criminal-scene are with the composite S TR amplification collection of illustrative plates of methyl alcohol replacement in conjunction with liquid II.
The toothbrush that Figure 11 leaves over for criminal-scene is with the composite S TR amplification collection of illustrative plates of propyl alcohol replacement in conjunction with liquid II.
The porcelain cup of the usefulness of drinking water that Figure 12 leaves over for criminal-scene is replaced composite S TR amplification collection of illustrative plates in conjunction with liquid II with Virahol.
Embodiment
Minim DNA on the belt hook that embodiment one extraction criminal-scene is left over.
(1) test kit is prepared
Preparation utilizes the test kit of magnetic bead extraction case trace sample DNA, comprising:
1mL silanization magnetic bead solution (0.1mg/ μ L), Wuhan Wawasye Technology Development Co., Ltd.'s listing product or Qiagen company buy;
100mL lysate, its component are the Tris-HCl damping fluid of pH=8.0;
100mL is in conjunction with liquid I, the PEG that its component volume ratio is 20-50% (molecular weight 8000);
20mL is in conjunction with liquid II, and its component is an ethanol;
100mL scavenging solution, its component are that volume ratio is 80% ethanolic soln.
(2) utilize test kit to extract the belt hook cast-off cells DNA that criminal-scene is left over, extraction step is as follows:
1) utilize case trace sample Adhesive-fetch device repeatedly the ixoderm bracelet repeatedly glued membrane is put into the 1.5mL centrifuge tube;
2) add 6 μ L Proteinase Ks (10mg/mL), 350 μ L lysates, whirlpool shook for 2 seconds, placed couveuse 56 degree and hatched 40 minutes;
3) centrifuge tube is taken off from couveuse, whirlpool shook for 2 seconds, and after placement couveuse 70-100 degree was hatched 8 minutes, 13000 left the heart 3 minutes, and supernatant is transferred to another 1.5mL centrifuge tube;
4) in conjunction with liquid II and 20 μ L magnetic bead suspensions, whirlpool shook for 2 seconds to adding 400 μ L in conjunction with liquid I, 80 μ L, placed room temperature 10 minutes;
5) with centrifuge tube as for magnetic force frame last 2 minute, inhale and abandon supernatant and stay magnetic bead, add 300 μ L scavenging solutions, whirlpool shook for 2 seconds, placed room temperature 2 minutes;
6) repeating step is 4 twice;
7) with centrifuge tube as for magnetic force frame last 2 minute, inhale and abandon supernatant and stay magnetic bead, room temperature was opened and placed to centrifuge tube 5 minutes;
8) add 30 μ L ultrapure waters, whirlpool shook for 2 seconds, placed couveuse 80 degree and hatched 10 minutes, and middle mixing magnetic bead 2 times places the magnetic force frame with centrifuge tube, and the gained supernatant is dna solution.
(3) pcr amplification and electrophoresis
Amplification kit is the sinofiler test kit of ABI company
Amplification program is as follows:
Electrophoresis equipment is 3100 type sequenators of ABI company
Last sample system: 2 μ L amplified productions+20 μ L methane amides (having added Liz)
Other steps: operate in strict accordance with 3100 type sequenator specification sheetss.
Fig. 1 leaves over the detected result of belt DNA for extracting this criminal-scene, 16 STR sites are complete, peak height meets case and detects the requirement that 16 STR sites are analyzed all more than 200, and the tool when the micro-case of extraction detects DNA that further illustrates this reagent and present method has great advantage.
Embodiment two control experiments
Is analog sample with 1 μ L through the blood of 60 times of physiological saline dilutions, replaces embodiment one cast-off cells glued membrane, and amplification and electrophoresis are with embodiment one, and it is as follows to design micro-sample extraction series controlled trial:
Controlled trial 1 extraction step 1) glued membrane is replaced by 1 μ L analog sample in, and other steps are constant;
Controlled trial 2 steps 2) do not add proteolytic enzyme in, other are with controlled trial 1;
Incubation temperature less than 70 degree in controlled trial 3 step 3), other are with test 1;
In conjunction with removing ethanol in the liquid system, other are with test 1 in controlled trial 4 step 4);
In conjunction with removing PEG in the liquid system, other are with test 1 in controlled trial 5 step 4);
Controlled trial 6 is through step 1), 2), 3), supernatant extracts DNA with Qiagen M48 magnetic bead kit;
Controlled trial 7 is not passed through step 1), 2), 3), extract DNA with M48 magnetic bead reagent;
Controlled trial 8 is through step 1), 2), 3, supernatant is used Microcon-100 purification column purifying concentration of DNA instead.
The dna solution that extracts through above experiment all carries out the multiplex PCR composite amplification with the ID test kit of ABI company with the system of 4 μ L template+6 μ LMix, and carries out electrophoresis detection with identical system and condition, gained result such as Fig. 2-and shown in Figure 9.Wherein, Fig. 2 is the result of serial controlled trial 1: 16 STR sites are complete, peak height is all more than 800, meet case and detect the requirement that 16 STR sites are analyzed, illustrate that cast-off cells DNA can efficiently discharge after bathing processing through protease digestion, temperature, efficiently is adsorbed on magnetic bead surfaces in PEG-ethanol articulated system.Fig. 3 is the result of serial controlled trial 2: the peak in CSF1PO site has been lost, the peak in D12S391 and D3S1350 site has height that the end is arranged, the amplification energy imbalance is obvious, D7S820 and D18S51 site are many peak, assorted peak is promptly arranged, and the somatotype that these all have a strong impact on detected result does not meet case and detects the requirement that 16 STR sites are analyzed, test explanation cast-off cells are without protease digestion, and DNA discharges not exclusively or fracture.Fig. 4 is the result of serial controlled trial 3: the small segment effect better all goes out as D8S1179 and peak, D19S433 site, the peak of sheet degree is too low, below 50 as D7S820, and assorted peak phenomenon such as D13S317 site are arranged, DNA in the PEG-ethanol system is poor in conjunction with magnetic bead efficient in explanation, controlled trial 2 and 3 explanation protease digestions, 70-100 degree temperature are bathed the cell high-efficient released dna, and dna structure is stable, is beneficial to magnetic bead to combine in the PEG-ethanol system.Fig. 5 is the result of serial controlled trial 4: 16 sites all do not go out, Fig. 6 is the result of serial controlled trial 5, site D8S1179, D5S818, vWA and D8S1043 have only been gone out, but the peak is below 100, D8S1043 has assorted peak phenomenon, do not meet case and detect the requirement that 16 STR sites are analyzed, must there be ethanol and PEG simultaneously in controlled trial 4 and the 5 description taken in conjunction liquid system, guarantee DNA and magnetic bead joint efficiency height, dna structure is complete relatively, the dna fragmentation that obtains to be of convenient length can satisfy case detect in the needs of 16 STR site composite amplifications.Fig. 7 is the result of serial controlled trial 6, Fig. 8 is the result of serial controlled trial 7, and the two result is more or less the same, and 16 sites all do not go out, peak height is below 200, and assorted peak is also many, influences result's interpretation, does not meet case and detects the requirement that 16 STR sites are analyzed, Fig. 9 is the result of serial controlled trial 8, similar with Fig. 6, only gone out the peak in 6 sites, ratio of peak is lower.Lose that the peak shows the dna structure destroyed or the DNA extraction amount is low, can not obtain capacity, be of convenient length and satisfy the DNA of 16 STR sites amplifications.Test 7 explanation guanidinesalt magnetic bead (M48 reagent) articulated systems possibility destructive dna structures or joint efficiency are low.Test 6,8 interpret sample are after protease digestion, 70-100 degree temperature are bathed the efficient release of DNA, the not only efficient adsorption of DNA of PEG-ethanol magnetic bead articulated system but also guaranteed that dna structure is stable is better than guanidinesalt magnetic bead (M 48 reagent) articulated system and guanidinesalt purification column articulated system (Microcon-100 reagent)---and these two kinds of systems are micro-sample DNA extraction common agents.To sum up the result of 8 serial controlled trials contrast as can be known, the dna solution that extracts with reagent of the present invention and method can obtain complete STR somatotype, and the STR peak height is all more than 200, and when cracking condition and articulated system change, all do not obtain complete STR somatotype, the peak-to-peak value that has gone out is substantially below 200.
Embodiment three changes methyl alcohol and propyl alcohol, Virahol in conjunction with the ethanol of liquid II
To change methyl alcohol and propyl alcohol, Virahol respectively in conjunction with the ethanol of liquid II among the embodiment one, extract the DNA on the porcelain cup of slippers, the toothbrush in the on-the-spot case, the usefulness of drinking water with paramagnetic particle method respectively;
80 μ L in extraction step 3 change into respectively 80 μ L methyl alcohol and propyl alcohol, the Virahol in conjunction with liquid II, and other are operated with embodiment one.
The slippers that Figure 10 leaves over for criminal-scene are with the composite S TR amplification collection of illustrative plates of methyl alcohol replacement in conjunction with liquid II; The toothbrush that Figure 11 leaves over for criminal-scene is with the composite S TR amplification collection of illustrative plates of propyl alcohol replacement in conjunction with liquid II; The porcelain cup of the usefulness of drinking water that Figure 12 leaves over for criminal-scene is replaced composite S TR in conjunction with liquid II with Virahol.After changing methyl alcohol, propyl alcohol and Virahol respectively in conjunction with the ethanol of liquid II as can be known by Figure 10, Figure 11, Figure 12, it is complete that the detected result of extracting the criminal-scene biological material through same procedure is 16 STR sites, peak height is all more than 200, meet case and detect the requirement that 16 STR sites are analyzed, illustrate and use methyl alcohol, propyl alcohol and Virahol replacement in conjunction with II in this extracting method and this test kit.
Reference:
1. hot army equality, the processing and the detection method research of the difficult sample product of medical science, criminal technique, 2001,6.
2. Wang Qin etc., DNA-STR somatotype check human body case trace sample 2 examples, criminal technique, 2004,5.
3. Wang Qin etc., the fluorescent mark STR somatotype of human body case trace sample, law and medical journal, 2004,11 (1).
4. open celebrating rosy clouds etc., extract trace sample DNA and crack murder case 1 example, Journal of Forensic Sciences, 2007,2,1 (23).
5. Wang Yu is strong etc., case trace sample minim DNA detection scheme pre-test on the clothing, criminal technique, 2007,2.
6. Zhang Zhi is superfine, the finishing of magnetic silica microballoon and the application in the Plant Genome nucleic acid purification thereof, analytical chemistry research report, 2007,1 (35).
7. the Cong Xian tinkling of pieces of jade etc. adopts the silicon dioxide microsphere method to reclaim the experimental study of DNA, Chinese Journal of Immunology, 2005,5.
Claims (10)
1. improve the method for cast-off cells DNA extraction efficient, it is characterized in that comprising the steps, the sample that comprises cast-off cells with protease treatment, the 70-100 degree is hatched treated sample then, centrifuging and taking sample supernatant, in PEG-ethanol binding buffer system, allow the DNA in the supernatant combine, separate and obtain DNA with magnetic bead.
2. the described method of claim 1 is characterized in that, sample was hatched 3～8 minutes at the 70-100 degree after 56 degree are hatched earlier again.
3. the described method of claim 1 is characterized in that, the concentration of PEG is 20-50%w/v in the PEG-ethanol binding buffer system.
4. the described method of claim 1, wherein ethanol can use methyl alcohol, propyl alcohol or Virahol to substitute.
5. the described method of claim 1 is characterized in that, described magnetic bead is handled through silanization.
6. test kit that is used to implement the described method of claim 1, it is characterized in that the reagent in this test kit comprises magnetic bead suspension, lysate, in conjunction with liquid I, in conjunction with liquid II and scavenging solution, wherein, lysate consists of the Tris-HCl of pH=8.0, the EDTA of pH=8.0,50-80mmol/L; Consist of the PEG of 20-50%w/v, the MW=8000 of PEG in conjunction with liquid I; II consists of ethanol in conjunction with liquid; Scavenging solution consists of the ethanolic soln of 50-80%v/v.
7. the described test kit of claim 6 is characterized in that each reagent is formed and had both comprised and comprise its diluent again for its concentrated solution.
8. the described test kit of claim 6, it is characterized in that in conjunction with liquid I, can be hybrid packed in conjunction with liquid II at a reagent bottle.
9. the described test kit of claim 6 is characterized in that in conjunction with liquid II be methyl alcohol, propyl alcohol or Virahol.
10. method of utilizing the described method of claim 1 to extract case trace sample DNA is characterized in that carrying out according to the following steps:
The carrier that may contain cast-off cells that 1) will shift places centrifuge tube, and adds lysate and proteolytic enzyme, and whirlpool concussion mixing, centrifuge tube place on the couveuse 56 degree to hatch 40～60 minutes;
2) centrifuge tube whirlpool concussion mixing places after the 70-100 degree is hatched 3～8 minutes on the couveuse, and 13000 left the heart 3 minutes;
3) shift supernatant, add the magnetic bead suspension, in conjunction with liquid I with in conjunction with liquid II, the piping and druming mixing forms the suspension liquid of solid-liquid homodisperse;
4) the magnetic force frame magnetic bead that will adsorb DNA is isolated from the suspension liquid of solid-liquid homodisperse;
5) scavenging solution washing is adsorbed with magnetic bead 2-3 time of DNA, with the impurity flush away on the magnetic bead;
6) DNA on the ultrapure water desorption magnetic bead obtains pure spissated dna solution.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201110148869.7A CN102229927B (en) | 2011-06-03 | 2011-06-03 | Improve method and the reagent of rate of extraction of DNAs of castoff cells in case trace sample |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201110148869.7A CN102229927B (en) | 2011-06-03 | 2011-06-03 | Improve method and the reagent of rate of extraction of DNAs of castoff cells in case trace sample |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102229927A true CN102229927A (en) | 2011-11-02 |
| CN102229927B CN102229927B (en) | 2016-03-23 |
Family
ID=44842532
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201110148869.7A Active CN102229927B (en) | 2011-06-03 | 2011-06-03 | Improve method and the reagent of rate of extraction of DNAs of castoff cells in case trace sample |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102229927B (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103622701A (en) * | 2012-08-23 | 2014-03-12 | 武汉哇哇噻纳技术开发有限公司 | Method for showing fingerprints through silanization nanometer magnetic powder |
| CN104830833A (en) * | 2015-03-23 | 2015-08-12 | 上海惠文生物技术有限公司 | Method for extracting DNA (deoxyribonucleic acid) by virtue of combination of magnetic bead method and centrifuge shield |
| CN106701737A (en) * | 2015-11-17 | 2017-05-24 | 安诺优达基因科技(北京)有限公司 | High-efficiency DNA purified magnetic bead reagent with fragment selective purification capacity |
| CN108893465A (en) * | 2018-07-25 | 2018-11-27 | 古蔺县公安局 | Magnetic bead is used to show the purposes and impression of the hand trace DNA extraction method of impression of the hand trace |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1995013368A1 (en) * | 1993-11-11 | 1995-05-18 | Medinnova Sf | Isolation of nucleic acid |
| US20030215845A1 (en) * | 2002-02-19 | 2003-11-20 | Bille Todd William | Selective extraction of DNA from groups of cells |
| CN101397588A (en) * | 2008-11-06 | 2009-04-01 | 昆明理工大学 | Reagent and kit for acquisition and recovery trace amount DNA and use method |
| CN101613697A (en) * | 2009-08-05 | 2009-12-30 | 公安部物证鉴定中心 | A kind of method of extracting purify DNA |
| CN102220310A (en) * | 2009-08-05 | 2011-10-19 | 公安部物证鉴定中心 | Method for extracting and purifying spittle DNA |
-
2011
- 2011-06-03 CN CN201110148869.7A patent/CN102229927B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1995013368A1 (en) * | 1993-11-11 | 1995-05-18 | Medinnova Sf | Isolation of nucleic acid |
| US20030215845A1 (en) * | 2002-02-19 | 2003-11-20 | Bille Todd William | Selective extraction of DNA from groups of cells |
| CN101397588A (en) * | 2008-11-06 | 2009-04-01 | 昆明理工大学 | Reagent and kit for acquisition and recovery trace amount DNA and use method |
| CN101613697A (en) * | 2009-08-05 | 2009-12-30 | 公安部物证鉴定中心 | A kind of method of extracting purify DNA |
| CN102220310A (en) * | 2009-08-05 | 2011-10-19 | 公安部物证鉴定中心 | Method for extracting and purifying spittle DNA |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103622701A (en) * | 2012-08-23 | 2014-03-12 | 武汉哇哇噻纳技术开发有限公司 | Method for showing fingerprints through silanization nanometer magnetic powder |
| CN103622701B (en) * | 2012-08-23 | 2018-08-10 | 聂棱 | Silanization nano-magnetic powder shows fingerprint method |
| CN104830833A (en) * | 2015-03-23 | 2015-08-12 | 上海惠文生物技术有限公司 | Method for extracting DNA (deoxyribonucleic acid) by virtue of combination of magnetic bead method and centrifuge shield |
| CN106701737A (en) * | 2015-11-17 | 2017-05-24 | 安诺优达基因科技(北京)有限公司 | High-efficiency DNA purified magnetic bead reagent with fragment selective purification capacity |
| CN106701737B (en) * | 2015-11-17 | 2020-07-31 | 安诺优达基因科技(北京)有限公司 | High-efficiency DNA purification magnetic bead reagent with fragment selective purification capability |
| CN108893465A (en) * | 2018-07-25 | 2018-11-27 | 古蔺县公安局 | Magnetic bead is used to show the purposes and impression of the hand trace DNA extraction method of impression of the hand trace |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102229927B (en) | 2016-03-23 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP5876481B2 (en) | Ultrafast nucleic acid purification method | |
| CN101935646B (en) | Kit and method for extracting DNA from micro samples | |
| CN102115711B (en) | Micro-flow control chip and nucleic acid extracting and purifying method | |
| CN108220125B (en) | Nucleic acid extraction device | |
| CN105349532A (en) | Method and kit for extracting free nucleic acid by using paramagnetic particle method | |
| Riordan et al. | Removal of uranium from solution using residual brewery yeast: combined biosorption and precipitation | |
| ATE452626T1 (en) | METHODS AND MATERIALS FOR DETECTING GENETIC MATERIAL | |
| JP2011530084A (en) | Methods and systems for selective isolation of target biological molecules in a universal system | |
| CN105734045B (en) | A method of the quick multi-pass amount based on micro-fluidic chip extracts blood sample DNA | |
| CA2484062A1 (en) | Dna purification and recovery from high particulate and solids samples | |
| CN109385418A (en) | A kind of method extracted for virus in animal specimen/bacterial nucleic acid and reagent | |
| CN105524917A (en) | Kit for extracting blood genome DNA based on magnetic bead method and use method for kit | |
| JP2011525806A (en) | Nucleic acid extraction method | |
| CN102229927B (en) | Improve method and the reagent of rate of extraction of DNAs of castoff cells in case trace sample | |
| WO2004072229A3 (en) | Chemical treatment of biological samples for nucleic acid extraction and kits therefor | |
| CN103710338A (en) | Kit for extracting DNA (deoxyribonucleic acid) from white cells in human whole blood | |
| CN109022420A (en) | A kind of DNA extraction method based on magnetic bead, lysate and kit | |
| CN108977437A (en) | A kind of kit and its method extracting nucleic acid from serum plasma sample | |
| McLamb et al. | Comparison of the M‐Vac® wet‐vacuum‐based collection method to a wet‐swabbing method for DNA recovery on diluted bloodstained substrates | |
| Laurin et al. | New incompatibilities uncovered using the Promega DNA IQ™ chemistry | |
| CN101748116A (en) | Method for extracting large numbers of DNA with magnetic particulate | |
| CN108410861A (en) | A kind of extracting method and kit of hide middle ancient times DNA | |
| CN110846308A (en) | Method for extracting DNA from hair | |
| EP1903110B1 (en) | Device for recovering nucleic acid and method for recovering nucleic acid | |
| CN102965271B (en) | Device for quickly extracting pine wood nematode nucleic acid |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| C41 | Transfer of patent application or patent right or utility model | ||
| GR01 | Patent grant | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20160301 Address after: 100080 Institute of physics, Chinese Academy of Sciences, South Third Street, middle pass, Haidian District, Beijing Applicant after: Nie Leng Address before: 177 No. 432100 Xiaogan city of Hubei province Huaiyin Avenue Applicant before: Wuhan Wawa Saina Technology Development Co.,Ltd. |
|
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20190117 Address after: Room 1702, Dantaihu Building (Wulu Science Park), No. 9 East Taihu Road, Wuzhong District, Suzhou City, Jiangsu Province Patentee after: Suzhou Bairan Gene Technology Co., Ltd. Address before: 100080 Institute of Physics, Chinese Academy of Sciences, South Third Street, Zhongguan, Haidian District, Beijing Patentee before: Nie Leng |
|
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20210528 Address after: 518110 b303, building 1, Yinxing Zhijie phase II, No. 1301-76, sightseeing Road, Xinlan community, Guanlan street, Longhua District, Shenzhen City, Guangdong Province Patentee after: Shenzhen Boyue Gene Technology Co.,Ltd. Address before: Room 1702, Dantaihu Building (Wulu Science Park), No. 9 East Taihu Road, Wuzhong District, Suzhou City, Jiangsu Province Patentee before: Suzhou Bairan Gene Technology Co.,Ltd. |