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

Abstract

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.

Keywords:

• Bentonite
• cDNA synthesis
• Jatropha curcas
• RNA isolation

Acknowledgements

CSIR-CSMCRI Communication No. as provided by BDIM is 224. We thank Dr Pushpito K. Ghosh for his constant support and encouragement while undertaking this work; and the anonymous reviewer. The financial assistance from Department of Science and Technology (DST) and Council of Scientific and Industrial Research (CSIR), New Delhi, India is duly acknowledged. Mitali Dabi is thankful to the Academy of Scientific & Innovative Research (AcSIR) for enrolment in the PhD course.

Disclosure statement

No potential conflict of interest was reported by the authors.

Additional information

Funding

The financial assistance from Department of Science and Technology (DST) and Council of Scientific and Industrial Research (CSIR), New Delhi, India is duly acknowledged.