Electronic Journal of Plant Breeding

Open Access
SCOPUS
  • Year: 2019
  • Volume: 10
  • Issue: 1

An improved method for rapid isolation of DNA and RNA from leaves, flowers and roots of blackgram [Vigna mungo (L.) Hepper] for detection of begomovirus infection and RT-PCR.

1Homi Bhabha National Institute, Training School Complex, Anushaktinagar, Mumbai-400094, India

Nuclear Agriculture and Biotechnology Division, Bhabha Atomic Research Centre, Trombay, Mumbai-400085, India

Abstract

Blackgram is an important pulse crop grown for its protein rich seeds. Its tissues are rich in polyphenols which hampered pure nucleic acid extraction and thus use of molecular techniques in breeding programme. In this study, a well established DNA extraction method for legumes, Gem-CTAB method has been modified to isolate PCR compatible DNA from blackgram in a quick and economical way from a very small tissue. The amplification rate of DNA extracted through modified Gem-CTAB method was found to be high compared to Gem-CTAB method for begomovirus specific genes. While for RNA isolation, among different protocols, column based kit method followed by lithium chloride precipitation yielded high quality RNA for leaves compatible with RT-PCR. For flowers and roots, kit method alone or with its modifications yielded pure RNA. The above modified nucleic acid extraction method yielded PCR compatible DNA and RNA from phenolics rich blackgram tissues.

Keywords

DNA, RNA, Blackgram, Modified Gem-CTAB, TRIzol reagent