Modified method of DNA isolation and optimization of PCR protocol for RAPD analysis of endangered medicinal plant Nardostachys jatamansi D C

Abstract

An important but limiting step in any molecular biological work is the reliable method of DNA isolation and suitable protocol for PCR analysis. Due to abundance of phenols, polysaccharides, terpenoids and other secondary metabolites in the rhizome of Jatamansi, it is a major problem to get high quality DNA. To isolate quality DNAfor PCR analysis modified protocol developed in our lab to overcome such serious problems. We tried to optimize the concentration of PVP, â-mercaptoethanol and NaCl. Upon gel documentation of isolated DNA by modified method evinced single discrete band of genomic DNA and yielded significantly superior, 30-50 µg/g DNA from dry rhizome. The composition of PCR master mix was also modified for RAPD analysis which includes changing the concentration of MgCl and observed a good amplification. Reproducible amplifiable products were 2 observed in all accessions.