A modified protocol for rapid DNA isolation from cotton (Gossypium spp.)

MethodsX
Qasim AliTayyab Husnain

Abstract

Extraction of high-quality DNA from Gossypium (Cotton) species is notoriously difficult due to high contents of polysaccharides, quinones and polyphenols other secondary metabolites. Here, we describe a simple, rapid and modified procedure for high-quality DNA extraction from cotton, which is amenable for downstream analyses. In contrast to other CTAB methods, the described procedure is rapid, omits the use of liquid nitrogen, phenol, CsCl gradient ultracentrifugation, uses inexpensive and less hazardous reagents, and requires only ordinary laboratory equipment. The procedure employed the high concentration of NaCl and use of PVP-10 to rid the problems associated with polysaccharides and polyphenols, respectively. The average yield was approximately 10-15 μg of good quality DNA from 100 mg of tissue weight, which is adequate for projects, like genetic mapping or marker-assisted plant breeding. This protocol can be performed in as little as 3 h and may be adapted to high-throughput DNA isolation. •Buffers A and B were redesigned from Paterson et al. (1993) and Porebski et al. (1997), respectively.•Ribonuclease A was added before chloroform extraction.•A simple, rapid and inexpensive DNA extraction method is described.

Citations

Nov 9, 2019·Journal of Animal Physiology and Animal Nutrition·Ahmad Ali ShahidTayyab Husnain
Sep 29, 2020·Food and Chemical Toxicology : an International Journal Published for the British Industrial Biological Research Association·Qasim AliAbdul Qayyum Rao
Nov 29, 2021·Molecular Biology Reports·Mohsin ShadAhmad Ali Shahid

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Methods Mentioned

BETA
PCR
electrophoresis

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