An efficient protocol for genomic DNA isolation from field grown mature leaves of Penniseteum gluacum[(L) R. Br.]

作者：Chatrath Padaria J; Choudhary M; Tarafdar A

来源：Research Journal of Biotechnology, 2013, 8(8): 30-34.

• Pennisetum glaucum
• mature plant leaves
• genomic DNA isolation
• cetyl trimethylammonium bromide (CTAB)
• polyvinylpyrrolidone (PVP)
• beta-mercaptoethanol

Pearl millet, Penniseteum gluacum [(L) R. Br.] is a rich source of novel genes for tolerance to different abiotic stresses as salinity, drought and high temperature. To carry out work on genomics of P. gluacum for isolating abiotic stress responsive novel genes, high quantity of DNA in purified form is required. Field grown leaf samples of P. gluacum are rich in secondary metabolites and polysaccharides which greatly interfere during DNA isolation and purification process leading to poor quality and quantity of isolated DNA. Such DNA preparations hinder the downstream molecular biology studies as restriction, PCR amplification and cloning.
Here we report an improvement in quality and yield of the extracted DNA from leaves of field grown pearl millet plants through the modification in the cetyl trimethylammonium bromide (CTAB) DNA extraction buffer. This modified buffer included 2% CTAB, 2% polyvinylpyrrolidone and 0.5% beta-mercaptoethanol with rest of the constituents as same as CTAB method described by Doyle and Doyle(5). The isolated DNA proved amenable to downstream molecular studies like PCR amplification and restriction digestion.

• 出版日期2013-8