Sri Sumarsih, 2012
(2012)
Eksplorasi enzim pendegradasi sampah organik dengan pendekatan metagenomik.
UNIVERSITAS AIRLANGGA.
(Unpublished)
Abstract
The construction of metagenom library has been carried out in the vector pCC2FOS�I E. coli EPI300-TI R system. Metagenom DNA was extracted directly from humus soil using SDS-based lysis method. The DNA was analyzed by 1% agarose electrophoresis. Metagenom library was constructed in accordance with the procedures of CopyControl� Fosmid library production kit, in several stages: (I) fragmentation of genomic DNA, (2) fragment DNA end-repair, (3) selection and recovery of end-repaired DNA fragments, (4) ligation of fragment DNA into the vector CopyControl pCC2FOST1vl, (5) recombinant DNA packaging, (6) plating and selection of fosmid library. The transformants were grown overnight on LB agar containing 12.5 flg/ ml chLoramphenicoL The results showed that metagenom DNA was isolated from humus soil and cloned into fosmid pCC2FOS�I E. coli EPI300-TIR system, produced metagenom library of 420 clones. However, it was not yet known whether or not the clones that expressed the organic waste degrading enzymes. Therefore, in further research will be selected for organic waste degrading enzymes, characterization, sequencing, sequence analysis and its homology with the sequences data ofGeneBank.
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