An economical and efficient protocol for total RNA isolation from Jatropha curcas

Jatropha curcas is an important non feed crop, increasingly important as a biofuel crop. It is hardy and resistant to different stress conditions in the field. In the wastelands of Gujarat (India), it is being grown for land reclamation and for socio-economic benefits. The long coastline in this state also promotes the growth of a large number of halophytes. Exploiting the genetic resource of Jatropha and halophytes for drought and salt-induced gene is an important area of research. For the isolation of genes and to study the molecular mechanism a good qualitative and quantitative RNA is a prerequisite. Jatropha leaves have latex, and therefore isolating RNA using guanidine thiocyanate or cetyltrimethylammonium bromide did not yield desirable quality of RNA. This paper reports a very simple and economical protocol for the isolation of good quality RNA from Jatropha and a few halophytes. The sodium dodecyl sulphate was used as a detergent for lysis of plant cells in the extraction buffer along with bentonite, which inhibits the ribonuclease’s activity. The addition of water saturated phenol in mortar-pestle, during grinding, facilitated better homogenisation of the tissues. Absolute RNA precipitation was obtained with the help of 2-butoxyethanol. Further this RNA was used successfully in preparation of complementary DNA and subsequently used for gene isolation.