Isolation and identification of culturable soil microorganisms from Poring Hot Spring, Ranau, Malaysia.

Eighteen unknown microorganisms had been cultured and isolated from the soil sample in Poring Hot Spring, Ranau, Malaysia. These microorganisms were grown around 12 hours on LB agar culture medium at 50 °C. The microorganism was chosen depending on their visual morphology characteristics. After the pure culture of unknown microorganisms were obtained, the PCR reactions were carried out using forward primer, fD1and reverse primer rP2 to amplify its 16S rRNA. The 16S rRNA of eighteen of the unknown microorganisms was examined by agarose gel electrophoresis on an ethidium bromide-stained 0.8 % agarose gel. Only nine unknown microorganisms (Unknown Microorganism 1, Unknown Microorganism 2, Unknown Microorganism 3, Unknown Microorganism 4, Unknown Microorganism 6, Unknown Microorganism 10, Unknown Microorganism 11, Unknown Microorganism 12, and Unknown Microorganism 17) were able to amplify the 16S rRNA and the size of the 16S rRNA was estimated around 1,500 bp. All nine bands were recovered by using QlAquick Gel Extraction Kit (QIAGEN, USA) after electrophoresis. Only PCR products of 4 unknown microorganisms (Unknown Microorganism 1, Unknown Microorganism 2, Unknown Microorganism 3, and Unknown Microorganism 17) were recovered successfully. However, sequencing of these fragments was unsuccessfully due to the low concentration of template DNA.