Diversity of Cultivable Bacteria and Those with Anti-microbial Activities - umsbact01

The Luria Bertani medium (10 g/l tryptone, 5 g/l yeast extract and 5 g/l sodium chloride), and Nutrient medium (Peptone from meat 3.45 g/l, sodium chloride 5.1 g/l and peptone from casein 3.45 g/l) were diluted to 1/10 of its original concentration, and sterilized by autoclaving at 121�C at 15lb for 15 mins. Bacteria from liquid culture were diluted and plated onto Luria-Bertani and Nutrient Media containing 1.5% Bacto-Agar (Difco).

Single colony was picked and streaked onto fresh medium and the processed was repeated until a pure culture was obtained. Two ml of bacteria in liquid culture was used for genomic DNA extraction using the DNAeasy Kit (Qiagen) according to the instructions from the manufacturer. Universal primers 25f, 518r, 530f, 907r, 926f, 1114f, 1392r and 1525r were used in various combinations to amplify the 16S rDNA of bacteria.

Amplified 16S rDNA genes were sequenced and analyzed using the basic local alignment search tool (BLAST), a web-based software at ncbi. nlm.nih.gov/BLAST/. This project is funded by the Malaysian Antarctica Research Programme (MARP) and Intensifying Research in Priority Areas (IRPA) grants from Malaysia.