Lysis buffers for extracting nucleic acids

Patent metadata

Field	Value
Publication number	US-10894957-B2
Application number	US-201816013938-A
Country	US
Kind code	B2
Filing date	Jun 21, 2018
Priority date	Sep 16, 2009
Publication date	Jan 19, 2021
Grant date	Jan 19, 2021

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 kit for extracting nucleic acid from a solid sample comprising: a lysis buffer comprising 250 to 500 mM ethylene glycol tetraacetic acid and further comprising 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, and sodium dodecyl sulfate, 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, trypsin and combinations thereof.

2. The kit of claim 1 , wherein the solid sample is selected from a biological material and an adhesive material.

3. The kit of claim 1 , optionally comprising: a solution comprising a polymer and detergent; and magnetically attractable particles.

4. The kit of claim 3 , wherein the polymer comprises one or more of dextran, cellulose, cellulose derivatives, soluble starch, dextrin, cellodextrin, polyethylene glycol, heparin, glycogen, and combinations thereof.

Assignees

• Life Technologies Corp

Inventors

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

Classifications

• 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

• C12N15/1013

  by using magnetic beads · CPC title

• C12N1/06

  Lysis of microorganisms · CPC title

• C12Q1/6806

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

• C12N15/1006

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