Microwave extraction of total genomic DNA from barley grains for use in PCR

H S. Saini
Mervyn Shepherd, Southern Cross University
Robert J. Henry, Southern Cross University

Abstract

Routine analysis of DNA from numbers of plant samples using the polymerase chain reaction (PCR) require procedures for rapid DNA preparation with the minimum number of steps and a reduced possibility of contamination. A simple and rapid method of DNA extraction from half or whole barley seeds employing microwave treatment for 60 seconds had been developed. The resulting extracts suitably diluted (1:10 to 1:20) were used in PCR with a conserved 5S ribosomal gene primer in conjunction with species-specific primers to amplify the non-coding spacer regions. Provided the extraction conditions are accurately established, the procedure allows up to 20 extractions in 60 seconds. Tlte procedure is time and cost effective and eliminates several sources of possible contamination associated with many other isolation procedures.