Defence Institute of Bio-Energy Research (DIBER), Goraparao, PO Arjunpur, Haldwani-263139, Uttarakhand, India
*Email: madhulogin@yahoo.co.in
Online published on 29 March, 2016.
Although thereisnodearth ofpublishedprotocols for DNA extraction in wide range of species, most of them are lengthy, time consuming and require more number of chemicals hence, not cost-effective. Also the amount of sample required for DNA extraction is large, which makes it difficult in extracting DNA from limited tissues. We have developed a simple, rapid and resource saving DNA extraction protocol for limited amount of plant tissues. This protocol work efficiently in wide range of plant species tried andhence, can beusedin anyspeciesfromwhich onewishes to extract DNA. This is also applicable for frozen preserved tissues. The extracted DNA was tested for PCR analysis using molecular markers like RAPD, EST-SSR primers and gene specific primers for evaluation of transgenic lines in capsicum and pea and also for hybridity confirmation of in vitro rescued wide hybrids in chickpea. The DNA extracted from this protocol was also found suitable for multiplexing using different primers of different product size but same melting temperature. This demonstrates the usefulness of this protocol since, DNA from more than 100 samples can bemanuallyor mechanicallyextracted within a day, making it a promising alternative in wide range of PCR based applications.
DNA isolation, Extraction buffer, PCR applications, Multiplexing