WO2012043183A1 - Procédé de synthèse d'un acide nucléique cible dans les selles - Google Patents

Procédé de synthèse d'un acide nucléique cible dans les selles Download PDF

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WO2012043183A1
WO2012043183A1 PCT/JP2011/070473 JP2011070473W WO2012043183A1 WO 2012043183 A1 WO2012043183 A1 WO 2012043183A1 JP 2011070473 W JP2011070473 W JP 2011070473W WO 2012043183 A1 WO2012043183 A1 WO 2012043183A1
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nucleic acid
stool
target nucleic
synthesizing
reaction
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PCT/JP2011/070473
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Japanese (ja)
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長岡　智紀
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オリンパス株式会社
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
    • C12N15/1096—Processes for the isolation, preparation or purification of DNA or RNA cDNA Synthesis; Subtracted cDNA library construction, e.g. RT, RT-PCR
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
    • C12N15/1003—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6806—Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay

Definitions

  • the present invention relates to a method for highly accurately synthesizing a target nucleic acid to be analyzed in feces using a nucleic acid synthesis reaction.
  • Patent Document 1 and Non-Patent Document 1 include a method for examining colon cancer by detecting non-apoptotic DNA frequently observed in nucleic acids derived from cancer cells, in particular, Alu repeat region and alphoid repeat region, A method for examining colorectal cancer based on the difference in fragment length of cancer-related genes such as p53 has been disclosed.
  • nucleic acid such as nucleic acid derived from cancer cells in stool
  • feces contain a large amount of digest residue and bacteria
  • nucleic acids are very easily degraded.
  • the analysis accuracy is impaired by introducing foreign substances in the stool into the nucleic acid collected from the stool. For this reason, in order to obtain a more reliable nucleic acid analysis result, a method for recovering highly purified nucleic acid from feces while preventing decomposition or the like has been developed.
  • Patent Document 2 discloses a method of stabilizing the stool structure by cooling the stool to a temperature below the gel freezing point, separating cells from the stool in this state, and analyzing the DNA extracted therefrom. ing.
  • Non-Patent Document 2 discloses that after removing contaminants such as proteins from a stool sample, RNA is extracted using phenol and chaotropic salt, and the extracted RNA is further extracted.
  • a method of recovering by adsorbing on a silica-containing solid support is disclosed.
  • Patent Document 3 discloses a method for preparing a stool sample for analyzing an oncogene in stool. This is a method in which a stool sample is homogenized at a solvent ratio of at least 5 with respect to stool mass 1, and then DNA derived from mammalian cells is collected including bacterial DNA.
  • the collected stool is homogenized in the presence of an RNase inhibitor, RNA is directly extracted from the prepared suspension, and Cox-2 (cyclooxygenase-2), which is an oncogene.
  • a method for detecting a transcript of a gene is disclosed.
  • feces contain substances having an inhibitory action on nucleic acid synthesis reactions such as PCR (Polymerase Chain Reaction) such as bile acids and salts thereof (for example, see Non-Patent Document 3).
  • PCR Polymerase Chain Reaction
  • bile acids and salts thereof for example, see Non-Patent Document 3.
  • the average amount of feces excreted by an adult is about 200 to 400 g / day, but there is a report that 200 to 650 mg / day of bile acid is excreted in feces of healthy people. That is, when converted to 1 g of stool, a healthy person contains about 0.5 mg to 3.25 mg, and a patient contains 10 times as much bile acid.
  • JP 2005-514073 A Japanese National Patent Publication No. 11-511982 Special Table 2002-539765 Japanese Patent No. 4134047 WO2010 / 024251A1
  • iron chloride is known to inhibit nucleic acid synthesis reactions such as PCR. Further, iron chloride is metabolized by bacteria in feces and the like, and there is a possibility that reactions such as PCR may be inhibited by irons other than iron chloride.
  • the present invention is a method of synthesizing a target nucleic acid to be analyzed in stool using a nucleic acid synthesis reaction, and an object thereof is to improve reaction efficiency and obtain a highly reliable result.
  • the present inventor reacted in the presence of an iron chelator when the nucleic acid extracted / recovered from stool is used as a template for a nucleic acid synthesis reaction such as reverse transcription reaction or PCR. As a result, it was found that the reaction efficiency can be improved, and the present invention was completed.
  • the present invention has the following configuration.
  • (1) A method for synthesizing a target nucleic acid in stool, in which a nucleic acid synthesis reaction using a nucleic acid collected from stool as a template in a reaction solution containing an iron chelator, (2) The method for synthesizing a target nucleic acid in feces according to (1), wherein the iron chelator is tetrasodium 3-hydroxy-2,2′-iminodisuccinate, (3) The method for synthesizing a target nucleic acid in stool according to (1) or (2), wherein the reaction solution further contains EDTA, (4) The method for synthesizing a target nucleic acid in stool according to any one of (1) to (3), wherein the nucleic acid synthesis reaction is a reverse transcription reaction or a nucleic acid amplification reaction using a polymerase, (5) The nucleic acid collected from the stool is a nucleic acid collected from the extraction solution after the nucleic acid extraction solution is added to and mixed with the stool
  • Example 1 it is the graph which showed the relative value of the expression level (copy number) of Cox-2 gene in RNA collect
  • the nucleic acid synthesis reaction means a reaction for synthesizing a nucleic acid using a reverse transcription reaction by a reverse transcriptase (reverse transcriptase) or a base chain extension reaction by a polymerase or ligase.
  • a reverse transcriptase reverse transcriptase
  • a base chain extension reaction by a polymerase or ligase examples include PCR (polymerase chain reaction), real-time PCR, and SDA (Standard Displacement Amplification).
  • Examples of the base chain extension reaction by ligase include LCR (ligase chain reaction).
  • the method for synthesizing a target nucleic acid in stool of the present invention (hereinafter sometimes referred to as the synthesis method of the present invention) is a method of synthesizing a target nucleic acid in stool, using a nucleic acid collected from stool as a template
  • the nucleic acid synthesis reaction is performed in a reaction solution containing an iron chelator.
  • an inhibitory substance means a substance that acts in an inhibitory manner on an enzymatic reaction using a nucleic acid as a substrate.
  • the enzyme reaction is not particularly limited as long as it is an enzyme reaction using a nucleic acid as a substrate, and also includes a nucleic acid synthesis reaction such as reverse transcription reaction and PCR.
  • the reaction efficiency of the nucleic acid synthesis reaction can be improved by adding an iron chelating agent to the reaction solution.
  • the reason why such an effect is obtained is not clear, but it is presumed that the reaction efficiency can be increased as a result of reducing the influence of the inhibitor remaining in the nucleic acid collected from the stool.
  • the type of inhibitor whose inhibitory action is reduced by the iron chelator is not clear, but is thought to be iron ions or their inclusions.
  • the iron chelating agent used in the synthesis method of the present invention is not particularly limited as long as it is a compound capable of chelating iron ions, but is preferably an iron chelating agent having high specificity for iron ions.
  • an iron chelating agent with high affinity for other ions such as magnesium ion, it may chelate even the ions required for the enzyme activity used in the nucleic acid synthesis reaction. Pay attention to the amount.
  • iron chelating agent used in the synthesis method of the present invention include 3-hydroxy-2,2′-iminodisuccinic acid tetrasodium (HIDS), EDTA (ethylenediaminetetraacetic acid), deferoxamine, deferiprone, deferasirox. Etc.
  • HIDS is preferably used because of its high specificity to iron ions.
  • the iron chelating agent added to the reaction solution may be only one type, or two or more types may be used in combination.
  • a higher reaction efficiency improvement effect can be achieved by using HIDS and EDTA together in appropriate amounts.
  • the concentration of the iron chelating agent added to the reaction solution in the synthesis method of the present invention is not particularly limited as long as the concentration is sufficient to obtain the reaction efficiency improvement effect of the nucleic acid synthesis reaction. It can be determined as appropriate in consideration of the type of nucleic acid, the amount of nucleic acid added to the reaction solution, the method for recovering nucleic acid from stool, and the like.
  • the HIDS concentration in the reaction solution is preferably 5 mM or more, more preferably 5 mM to 1 M, and even more preferably 10 mM to 1 M.
  • the EDTA concentration in the reaction solution is preferably 10 mM to 0.1 M.
  • the HIDS content in the reaction solution is preferably 5 mM or more and the EDTA content is preferably 1 to 10 mM.
  • the nucleic acid used as a template for the nucleic acid synthesis reaction may be a nucleic acid recovered from stool, and may be a nucleic acid recovered from stool by any known method.
  • it may be a nucleic acid obtained by separating and recovering a cell containing a target nucleic acid from stool in advance, as in Patent Document 2 and the like, and extracted and purified from the recovered cell.
  • It may be a nucleic acid recovered directly from stool without going through a separation step. Since the effect of improving the reaction efficiency of the nucleic acid synthesis reaction of the present invention can be exhibited more remarkably, in the present invention, it is preferable to use a nucleic acid recovered directly from stool or a solid component thereof.
  • Examples of the method for directly recovering nucleic acid from stool or its solid component include a method of adding a nucleic acid extraction solution directly to stool and mixing it, and then recovering the nucleic acid eluted in the extraction solution.
  • the method involves nucleic acids of all biological species contained in the stool, mainly nucleic acids derived from animals excreting the stool, and bacteria such as intestinal resident bacteria without performing the separation operation of cells and contaminants.
  • the nucleic acid derived from the stool is simultaneously extracted and recovered.
  • the nucleic acid contained in the stool includes, in addition to animal-derived nucleic acids and bacteria-derived nucleic acids, food-derived nucleic acids ingested by the animals.
  • the extraction solution may be any solution that can denature proteins in solid components and elute nucleic acids into the extraction solution from cells such as mammalian cells and intestinal resident bacteria in the solid content.
  • the solution is not particularly limited, and any solution used in the technical field may be used.
  • a solution in which a compound usually used as a protein denaturant such as a chaotropic salt, an organic solvent, or a surfactant is added as an active ingredient to an appropriate solvent can be used as the extraction solution.
  • These active ingredients may be a combination of two or more.
  • the surfactant that can be an active ingredient of the extraction solution is preferably a nonionic surfactant.
  • the nonionic surfactant include Tween 80, CHAPS (3- [3-Colamidopropyldimethylammonio] -1-propanesulfonate), Triton X-100, Tween 20, and the like.
  • the concentration of the chaotropic salt and the surfactant is not particularly limited as long as the nucleic acid can be eluted from the solid component, and the mixing ratio of the stool amount (solid component amount) and the extraction solution, or the recovered amount is recovered. It can be determined appropriately in consideration of the nucleic acid synthesis method.
  • Phenol is preferable as the organic solvent that can be an active ingredient of the extraction solution.
  • Phenol may be neutral or acidic. When acidic phenol is used, RNA can be selectively extracted into the aqueous layer rather than DNA.
  • a solvent for adding these active ingredients to prepare an extraction solution for example, a phosphate buffer or a Tris buffer can be used.
  • a drug in which DNase is inactivated by high-pressure steam sterilization or the like is preferable, and a drug containing a protease such as proteinase K is more preferable.
  • a citrate buffer or the like can be used as the elution agent.
  • RNA is a substance that is very easily decomposed, so RNase such as guanidine thiocyanate or guanidine hydrochloride is used. It is preferable to use a buffer containing an inhibitor.
  • the solid component Before adding the extraction solution, other solutions such as an appropriate storage solution are added to the collected stool, and if the suspension is in the form of a suspension, the solid component is recovered from the suspension. You may add the solution for extraction to the collect
  • the recovery of the solid component from the suspension can be performed by a known solid-liquid separation process such as a centrifugal separation process or a filter filtration process.
  • the recovered solid component may be washed with an appropriate buffer and then the extraction solution may be added.
  • any conventionally known solution may be used, but a solution containing a nucleic acid stabilizer is preferable.
  • the nucleic acid stabilizer include water-soluble organic solvents, protease inhibitors, polycations, and high-concentration salts.
  • Recovery of the nucleic acid eluted in the extraction solution can be performed by a known method such as ethanol precipitation or cesium chloride ultracentrifugation. Further, after the nucleic acid eluted in the extraction solution is adsorbed on the inorganic support, the nucleic acid can be recovered by eluting the adsorbed nucleic acid from the inorganic support using a certain volume of solvent. .
  • the inorganic support for adsorbing nucleic acid a known inorganic support capable of adsorbing nucleic acid can be used.
  • the shape of the inorganic support is not particularly limited, and may be in the form of particles or a film.
  • the inorganic support examples include silica-containing particles (beads) such as silica gel, siliceous oxide, glass, and diatomaceous earth, and porous membranes such as nylon, polycarbonate, polyacrylate, and nitrocellulose.
  • Solvents for eluting adsorbed nucleic acids from inorganic supports are usually used to elute nucleic acids from these known inorganic supports in consideration of the type of nucleic acid to be recovered, the subsequent nucleic acid analysis method, and the like.
  • a solvent can be appropriately used.
  • the elution solvent is particularly preferably purified water.
  • the inorganic support on which the nucleic acid has been adsorbed is preferably washed with an appropriate washing buffer before the nucleic acid is eluted.
  • the denatured protein Before recovering the nucleic acid, the denatured protein may be removed from the extraction solution from which the nucleic acid has been eluted.
  • the quality of the recovered nucleic acid can be improved by removing the protein that has been denatured in advance before recovering the nucleic acid.
  • the protein can be removed from the extraction solution by a known method.
  • the denatured protein can be removed by precipitating the denatured protein by centrifugation and collecting only the supernatant.
  • centrifugation is performed, and the denatured protein is precipitated and only the supernatant is recovered. Protein can be removed.
  • nucleic acid recovery from feces can also be performed using a commercially available kit such as a nucleic acid extraction kit.
  • a nucleic acid synthesis reaction is performed in a reaction solution containing an iron chelator.
  • the nucleic acid synthesis reaction can be performed by a conventional method except that an iron chelator is added to the reaction solution.
  • the target nucleic acid in the present invention is a nucleic acid to be synthesized, and is particularly limited as long as it has a nucleotide sequence that has been clarified to the extent that it can be analyzed by a method used for analysis of normal nucleic acids such as PCR. It is not something.
  • the target nucleic acid in the present invention is preferably a nucleic acid derived from a mammalian cell, and more preferably an RNA derived from a mammalian cell (for example, mRNA or the like).
  • RNA expression level can also be detected.
  • mRNA expression analysis K-ras gene mutation analysis
  • DNA methylation analysis DNA methylation analysis
  • analyzes can be performed by methods known in the art.
  • a commercially available analysis kit such as a K-ras gene mutation analysis kit or a methylation detection kit may also be used.
  • nucleic acid recovered from feces since it is a nucleic acid recovered from feces, it is preferable to use a nucleic acid derived from gastrointestinal cells such as the large intestine, small intestine, stomach, etc. as a target nucleic acid, and a nucleic acid derived from a large intestine exfoliated cell as a target nucleic acid. Is more preferable.
  • a nucleic acid derived from a marker gene for neoplastic transformation (including cancer) or a marker gene for inflammatory gastrointestinal diseases is preferably used as a target nucleic acid
  • a nucleic acid derived from a marker gene for colon cancer is preferably used as a target nucleic acid.
  • the “gene-derived nucleic acid” means an expression product such as genomic DNA or mRNA of the gene.
  • the marker showing neoplastic conversion include known cancer markers such as Cox-2 gene, carcinoembryonic antigen (CEA), sialyl Tn antigen (STN), APC gene, p53 gene, K-ras gene, etc. The presence or absence of mutations.
  • markers that indicate inflammatory digestive tract diseases include Cox-2 gene-derived nucleic acids.
  • the Cox-2 gene-derived nucleic acid is also used as a marker indicating neoplastic transformation.
  • the reaction efficiency can be increased, and as a result, a stable reaction can be performed.
  • the synthesis method of the present invention it is possible to obtain a highly accurate and highly reliable result for the target nucleic acid in the nucleic acid recovered from stool containing a large amount of contaminants.
  • highly reliable results of gene expression analysis of stool-derived RNA can be obtained. The result thus obtained is preferably used for clinical examination.
  • a nucleic acid derived from a marker gene for colorectal cancer such as Cox-2 gene in feces can be synthesized with high accuracy.
  • the synthetic product thus obtained can be used for colorectal cancer detection. That is, information for clinical examination can be provided by using the synthesis method of the present invention.
  • the synthesis method of the present invention can be carried out more easily by making an iron chelating agent into a kit with a reagent used in a nucleic acid synthesis reaction.
  • a nucleic acid synthesis kit preferably includes HIDS or HIDS and EDTA as an iron chelator.
  • the reagent used for the nucleic acid synthesis (nucleic acid amplification) reaction include an enzyme (nucleic acid synthase), a buffer, a primer, and the like.
  • a product obtained by adding an iron chelator to a commercially available kit for reverse transcription reaction or PCR can be used as the nucleic acid synthesis kit of the present invention.
  • the kit for nucleic acid synthesis of the present invention may be provided with reagents used for recovering nucleic acid from stool.
  • reagents used for recovering nucleic acid from stool.
  • examples of such reagents include a storage solution added to feces, a nucleic acid extraction solution, an inorganic support, a solvent for elution from the inorganic support, and the like.
  • what added the iron chelating agent to the commercially available nucleic acid extraction kit can also be used as the nucleic acid synthesis kit of the present invention. In the present invention, it is preferable to dissolve the iron chelating agent in advance in the solvent for elution from the inorganic support, because the labor of mixing can be saved.
  • MKN45 cells used were cultured by a conventional method.
  • Example 1 Synthesis of nucleic acid derived from Cox-2 gene in RNA extracted from stool derived from pseudo-colon cancer patient
  • derived from Cox-2 gene in RNA extracted from stool sample derived from pseudo-colorectal cancer patient Nucleic acids were synthesized and detected.
  • a stool sample derived from a pseudo-colorectal cancer patient (a stool pseudo-sample collected from a colorectal cancer patient) contains MKN45 cells, a cultured cell line that highly expresses the Cox-2 gene, in the same manner as colorectal cancer cells. A mixture was used.
  • the obtained stool samples derived from pseudo colorectal cancer patients were dispensed into three 1.5 mL tubes of 100 ⁇ L each. To these three 1.5 mL tubes, 1 mL each of RLT buffer containing guanidine salt was added and stirred with Vortex. Centrifugation was performed after stirring, 300 ⁇ L of the supernatant was collected, and an equal amount of 70% ethanol was added and mixed. The obtained mixed solution is passed through an RNA recovery column of RNeasy midi kit (manufactured by Qiagen), and the RNA recovery column is subjected to washing operation and RNA elution operation according to the attached protocol, whereby 50 ⁇ L of total RNA is obtained. It was collected as a solution. The total RNA solution collected in three tubes was mixed to form one total RNA solution, the RNA concentration of the solution was measured, and the total RNA amount collected was quantified.
  • a reverse transcription reaction was performed in the presence of HIDS, EDTA, or BSA to synthesize cDNA.
  • a reverse transcription reaction was performed under the condition where none of HIDS, EDTA, and BSA was added.
  • Table 1 shows the concentrations of HIDS, EDTA, and BSA in the reaction solution of each sample.
  • a reaction solution for the reverse transcription reaction was prepared so that 1 ⁇ g of RNA was added to each reaction solution.
  • M-MLV (TaKaRa) was used as a reverse transcriptase.
  • Real-time PCR was performed using the obtained cDNA as a template, and the expression product (mRNA) of the Cox-2 gene was detected.
  • Cox-2 primer probe MIX (Catalog No: Hs00153133_m1) manufactured by Applied Biosystems was used. Specifically, 1 ⁇ L of each cDNA was dispensed into a 0.2 mL 96-well PCR plate. Thereafter, 8 ⁇ L of ultrapure water and 10 ⁇ L of nucleic acid amplification reagent “TaqMan GeneExpression Master Mix” (Applied Biosystems) were added to each well, and 1 ⁇ L of Cox-2 primer probe MIX (Applied Biosystems) was further added.
  • PCR reaction solution was prepared.
  • the PCR plate was placed in an ABI real-time PCR apparatus, treated at 95 ° C. for 10 minutes, and then subjected to 40 cycles of thermal cycling at 95 ° C. for 1 minute, 56.5 ° C. for 1 minute, and 72 ° C. for 1 minute. Further, PCR was performed while measuring the fluorescence intensity over time by treating at 72 ° C. for 7 minutes.
  • the measurement result of the fluorescence intensity was analyzed, and the expression level of the Cox-2 gene in each sample was examined.
  • the relative value of the expression level of other samples was calculated with the expression level of the control without addition of HIDS and the like (sample 17 in Table 1) being 1.
  • the calculation results are shown in the graph of FIG.
  • EDTA also has an effect of chelating trivalent iron ions (Fe 3+ ).
  • a target nucleic acid in stool can be synthesized and detected with high accuracy using a nucleic acid synthesis reaction, and thus can be used particularly in the field of clinical examinations.

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Abstract

La présente invention concerne un procédé qui permet de synthétiser un acide nucléique cible dans les selles en utilisant une réaction de synthèse d'acide nucléique, qui améliore le rendement de la réaction, et qui obtient un résultat extrêmement fiable. Le procédé de synthèse d'un acide nucléique cible dans les selles est caractérisé par l'exécution de la réaction de synthèse de l'acide nucléique, laquelle utilise un acide nucléique recueilli à partir des selles comme modèle, dans un liquide réactionnel contenant un chélateur du fer. Le chélateur du fer peut être du tétrasodium 3-hydroxy-2,2'-iminodisuccinate. La présente invention concerne en outre un procédé donnant des informations pour le contrôle du cancer du côlon en utilisant le procédé de synthèse d'un acide nucléique dans les selles. La présente invention concerne en outre un kit destiné à la synthèse d'un acide nucléique, qui présente un chélateur du fer, et qui est utilisé dans le procédé de synthèse d'un acide nucléique cible dans les selles.
PCT/JP2011/070473 2010-09-28 2011-09-08 Procédé de synthèse d'un acide nucléique cible dans les selles WO2012043183A1 (fr)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112204139A (zh) * 2018-04-02 2021-01-08 太阳基因组学公司 用于从样品中的一种或多种类型的微生物的不同群体中提取核酸分子的通用方法
US11959125B2 (en) 2016-09-15 2024-04-16 Sun Genomics, Inc. Universal method for extracting nucleic acid molecules from a diverse population of one or more types of microbes in a sample

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WO2010064634A1 (fr) * 2008-12-05 2010-06-10 オリンパス株式会社 Procédé pour préparer un échantillon fécal, solution pour préparer un échantillon fécal et kit de collecte de fèces

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US11959125B2 (en) 2016-09-15 2024-04-16 Sun Genomics, Inc. Universal method for extracting nucleic acid molecules from a diverse population of one or more types of microbes in a sample
CN112204139A (zh) * 2018-04-02 2021-01-08 太阳基因组学公司 用于从样品中的一种或多种类型的微生物的不同群体中提取核酸分子的通用方法
JP2021520191A (ja) * 2018-04-02 2021-08-19 サン ジェノミクス インコーポレイテッド 試料中の１つ以上の種類の微生物の多様な集団から核酸分子を抽出するための万能な方法

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