Bacterial Community Analysis of Gedongsongo Hot Spring : Denaturing Gradient Gel Electrophoresis

Abstract

The bacterial communities from one of hot spring at Gedongsongo (WGS2) Ambarawa, Central Java, Indonesia; was investigated by molecular analysis based on the 16S rRNA gene. Two minimal media, MM1 and MM2 were used for growth of aerobic microbial communities. Cultures media were combined by filtration through 0.2-µm-pore-size filter for total genomic DNA extraction. The DNA that was exctracted both from cells of filtration and cultures have been well characterized as microbial chromosomal DNA and used as PCR template. Partial 16S rRNA gene sequences were PCR amplified using one primer set. One primer complements a region conserved among members of the domain Bacteria (Eschericia coli positions 1055 to 1070. The other primer is based on a universally conserved region (E.coli positions 1392 to 1406 and incorporates a 40-base GC clamp. These primers amplified a 323-bp section of the 16S rRNA genes. The amplicons were separated by denaturing gradient gel electrophoresis (DGGE) for community analysis. The DGGE profiles showed that there were three distinct bands, but only two of them that represent the predominant bacteria.