A simplified method for extraction of high quality genomic DNA from Jatropha curcas for genetic diversity and molecular marker studies

摘要

A simple and efficient protocol for the extraction of high quality genomic DNA from different tissues, including callus generated from leaves of Jatropha curcas has been developed. The important steps in this protocol include (a) use of 3.5 M NaCl in extraction buffer; (b) 2.0 M NaCl (final concentration) during precipitation; (c) Tris saturated phenol in place of phenol: chloroform: isoamyl alcohol at purification phase; (d) 80% ethanol for DNA precipitation, and (e) performing all the steps at RT. The DNA thus extracted from the leaves had 1.81 +/- 0.063, OD at A(260/280) and the yield was 120 to 140 mu g/g of material. The extracted DNA was found suitable for restriction digestion, ligation and PCR amplification. It was also used for DNA fingerprinting techniques, RAPD and AFLP, for development of molecular markers and studies on genetic diversity.