Cellulolytic Bacteria Degradation DNA Isolation from Acid Soil Molecularly

Abstract

AbstractCellulolytic degradation bacteria are microorganisms that are abundant in nature which can break down raw fibers of organic waste into compost. The purpose of this study was to obtain superior cellulolytic bacteria to degrade organic waste into compost and obtain permanent species and strain. The method used is DNA isolation by culturing bacteria with Luria Bertani (LB) media, quality and quantity analysis using DNA electrophoresis isolation technique visualized by electrophoresis technique followed by testing with Polymerase Chain Reaction (PCR) technique. Isolated DNA was amplified using a primer combination designed from the 16S rRNA gene sequence. The primary combinations used are 27F (5 '-AGAGTTTGATCMTGGCTCAG-3') and 1525R (5 '-AAGGAGGTGWTCC-ARCC-3'). Analysis of sequence data is compared with gene sequences that have been deposited into the public database in silico using the BLAST program conducted online on the NCBI website (National Center of Biotechnology Information). The results obtained by visualization of electrophoresis from isolation of DNA 6 isolates of cellulolytic bacteria M = λ DNA 50 ng / µl; 1 = KM25, 2 = SR75, 3 = JM, 4 = U6,5 = G8,6 = K13. DNA concentrations obtained from 6 isolates were quite high, ranging from 34.4 to 69.9 ng / µL. The results of the 6th PCR amplification were 6 isolates namely KM25, SR75, JM, G-8, KM13, AND U-6. In general, the six selected cellulolytic bacterial isolates belong to the genus Bacillus. Phylogenetic analysis using Boostrap 1000 x. The six selected bacterial isolates analyzed separated and formed two main clusters: the first cluster which included Bacillus thuringensis and Bacillus cereus and the Bacillus Subtilis cluster. The conclusion of the 6 isolates sequencing was KM13 (Bacillus thuringensis), G8 (Bacillus cereus), Sr75 (Bacillus cereus), JM (Bacillus subtilis), KM25 (Bacillus subtilis), U6 (Bacillus subtilis).

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