Lysis buffers for extracting nucleic acids

Patent metadata

Field	Value
Publication number	US-9447409-B2
Application number	US-88219410-A
Country	US
Kind code	B2
Filing date	Sep 14, 2010
Priority date	Sep 16, 2009
Publication date	Sep 20, 2016
Grant date	Sep 20, 2016

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.

First claim

What is claimed is:

1. A composition comprising 250 to 500 mM ethylene glycol tetraacetic acid and each of the following: a detergent, a reducing agent, and an enzyme, wherein the detergent is selected from one or more of N-lauroyl sarcosine, sodium deoxycholate, cetyltrimethylammonium bromide, dodecyl β-D-maltoside, nonanoyl-N-methylglucamide, polyethylene glycol p-(1,1,3,3-tetramethylbutyl)-phenyl ether, sodium dodecyl sulfate, and combinations thereof, wherein the reducing agent comprises one or more of tris(2-carboxyethyl)phosphine, dithioerythritol, and dithiothreitol, and wherein the enzyme is selected from caspase, chymotrypsin, pepsin, proteinase K, thrombin, Staphylococcus V8 protease, pronase, papain, Bacillus sp. E1A protease, and trypsin and combinations thereof.

2. The composition of claim 1 , wherein the detergent is N-lauroyl sarcosine, the reducing agent is dithiothreitol, and the enzyme is proteinase K.

3. A method of making a product, wherein the product comprises a nucleic acid, comprising:
a) incubating a solid in a solution;
b) extracting a supernatant from the incubated solid in the solution, wherein the supernatant comprises the nucleic acid extracted from the solid, wherein the solution comprises 250 to 500 mM ethylene glycol tetraacetic acid and each of the following: a detergent, a reducing agent, and an enzyme, wherein the detergent is selected from one or more of N-lauroyl sarcosine, sodium deoxycholate, cetyltrimethylammonium bromide, dodecyl β-D-maltoside, nonanoyl-N-methylglucamide, polyethylene glycol p-(1,1,3,3-tetramethylbutyl)-phenyl ether, sodium dodecyl sulfate, and combinations thereof, wherein the reducing agent comprises one or more of tris(2-carboxyethyl)phosphine, dithioerythritol, and dithiothreitol, and wherein the enzyme comprises one or more of caspase, chymotrypsin, pepsin, proteinase K, thrombin, Staphylococcus V8 protease, pronase, papain, Bacillus sp. E1A protease, and trypsin and combinations thereof.

4. A method of identification comprising:
(a) lysing a sample in a lysis solution, forming a lysate comprising a nucleic acid from the sample, wherein the lysis solution comprises 250 to 500 mM ethylene glycol tetraacetic acid and each of the following: a detergent, a reducing agent, and an enzyme, wherein the detergent is selected from one or more of N-lauroyl sarcosine, sodium deoxycholate, cetyltrimethylammonium bromide, dodecyl β-D-maltoside, nonanoyl-N-methylglucamide, polyethylene glycol p-(1,1,3,3-tetramethylbutyl)-phenyl ether, sodium dodecyl sulfate, and combinations thereof, wherein the reducing agent comprises one or more of tris(2-carboxyethyl)phosphine, dithioerythritol, and dithiothreitol, and wherein the enzyme comprises one or more of caspase, chymotrypsin, pepsin, proteinase K, thrombin, Staphylococcus V8 protease, pronase, papain, Bacillus sp. E1A protease, and trypsin and combinations thereof;
(b) extracting the lysate, wherein the nucleic acid is in the lysate;
(c) analyzing the nucleic acid; and
(d) identifying the sample based on the nucleic acid analysis.

5. The method according to claim 4 , wherein the sample is a fingerprint tape lift.

6. The method of claim 5 , further comprising comparing the fingerprint to a collection of fingerprints.

Assignees

• Stray James
• Liu Jason Yingjie
• Brevnov Maxim
• Shewale Jaiprakash
• Holt Allison
• Life Technologies Corp

Inventors

• Stray James
• Liu Jason Yingjie
• Brevnov Maxim
• Shewale Jaiprakash
• Holt Allison

Classifications

• C12N15/1013

  by using magnetic beads · CPC title

• C12N15/1003 Primary

  Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor · CPC title

• C12N1/06

  Lysis of microorganisms · CPC title

• C12N15/1006 Primary

  by means of a solid support carrier, e.g. particles, polymers · CPC title

• C12Q1/6806

  Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay (C12Q1/6804 takes precedence) · CPC title