Isolation and PCR Amplification of Genomic DNA From Traded Seeds Of Nutmeg (M. Fragrans)

Abstract

An efficient protocol for the isolation of high molecular weight DNA from powder of traded nutmeg seeds (Myristica fragrans) is described which will aid in protecting the intellectual property rights of those who trade nutmeg in the world market. The method involves a modified 3 % CTAB (hexadecyltrimethylammonium bromide) procedure with 2M NaCl, 0.3 per cent mercaptoethanol and 1.5% polyvinylpyrrolidone. The yield of the DNA obtained from the samples varied from 4 to 6 ug/g tissue. The isolated DNA was subjected to Polymerase Chain Reaction (PCR) amplification using a random decamer primer 5 GGACCCTTAC3 and showed distinct and reproducible band patterns. The protocol has trade implications as it will help in the PCR characterization of traded nutmeg seeds and powders from different regions and thus help in tracing the geographical origin of the commodity.

Keywords: Nutmeg, Myristica fragrans, RAPD, PCR, DNA, spice, powder, recalcitrant