LYSIS BUFFER AND METHODS FOR EXTRACTION OF DNA FROM PLANT MATERIAL

摘要

The present invention is generally directed to a lysis buffer for extraction of DNA from plant material and improved methods for extraction of DNA from plant material utilizing the novel lysis buffer. Advantageously, the lysis buffer of the present invention is suitable for use in connection with simpler analysis methods, while still providing suitable DNA yields and purities for analysis.

主权项

1. A cell lysis buffer composition, the composition comprising:
(a) a buffering component selected from the group consisting of tris(hydroxymethyl)aminomethane (Tris), 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), 3-(N-morpholino)propanesulfonic acid (MOPS), sodium dihydrogen phosphate (NaH2PO4), disodium hydrogen phosphate (Na2HPO4), and combinations thereof; (b) a mineral salt selected from the group consisting of sodium chloride (NaCl), potassium chloride (KCl), diammonium sulfate (NH4SO4), and combinations thereof; (c) a metal chelating agent selected from the group consisting of ethylenediaminetetraacetic acid (EDTA), ethylene glycol tetraacetic acid (EGTA), and combinations thereof; and (d) a surfactant selected from the group consisting of sodium dodecyl sulfate (SDS), nonyl phenoxypolyoxylethanol (NP-40), polyethylene glycol p-(1,1,3,3-tetramethylbutyl)-phenyl ether (triton-X), polyoxyethylene (20) sorbitan monooleate (Tween-20), and combinations thereof, the composition further comprising one or more of the following components: (A) an alkali metal hydroxide selected from the group consisting of sodium hydroxide, potassium hydroxide, and combinations thereof; and/or (B) a precipitant selected from the group consisting of glycerol, dimethyl sulfoxide (DMSO), acetonitrile (ACN), bovine serum albumin (BSA), proteinase K, acetate salts, and combinations thereof; and/or (C) a water-soluble polymer comprising polyvinylpyrrolidone.