RAPID AND EFFICIENT GENERATION OF PCR TEMPLATES FROM ESCHERICHIA COLI, SACCHAROMYCES CEREVISIAE AND ORYZA SATIVA USING A MICROWAVE AND BY BOILING

Abstract

Simple methods for the preparation of polymerase chain reaction (PCR) templates from Escherichia coli, Saccharomyces cerevisiae and Oryza sativa using a microwave and by boiling are described without the need for any special chemical reagent such as a detergent. The microwave and boiling methods were robust for the preparation of both plasmid and genomic templates of E. coli, as was the boiling approach for S. cerevisiae DNA. The microwave procedure was only suitable for dry yeast colonies with fair efficiency as absence of water (H2O) was critical for a successful PCR. Oryza sativa genomic DNA was amplified with both approaches with satisfactory efficiency. Escherichia coli plasmids can be prepared using a microwave with a yield equivalent to 15–20% of an alkaline lysis mini‐prep. PCR templates were also efficiently generated from Pichia pastoris and Ganoderma lucidum by using the boiling method.

Isolation of novel sequences targeting highly variable viral protein hemagglutinin

“…Cells were plated onto LB/ampicillin plates. PCR was performed with primers 4 and 9 after microwave treatment of toothpick touched colonies [6] , and 1550 bp PCR amplicons were visualized on agarose gel ( Fig. 3 ).…”