IJSTR >> Volume 11 - Issue 01, January 2022 Edition

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AUTHOR(S)

Michael G. Garlan

KEYWORDS

gDNA, Polymerase Chain Reaction, Gel Electrophoresis, Spectrophotometry, Staphylococcus aureus, elution, buffer

ABSTRACT

This paper aimed to model purification, amplification and visualization a microbial genomic DNA (gDNA) eluted from Staphylococcus aureus prepared overnight at 37˚C and to compare the amplified gDNA with the standard DNA ladder 1 kb as standardized procedure for student research. The purity of the gDNA was also determined using spectrophotometric analysis with the A260/A280 ratio of 2.0 as the highest and 1.9 as the lowest which indicated the gDNA eluted was within the purity range. The genomic DNA with 4 replicates (A1, A2, A3 and A4) were amplified through polymerase chain reaction (PCR) machine for 1 hour using PCR master mix of 5 µL PCR buffer, 1 µL forward primer, 1 µL reverse primer, 12.8 µL SnpH2O and 0.2 µL Taq polymerase. The amplified DNA as well as the gDNA quality were then determined by loading the EtBr-stained gel agarose wells with DNA samples including the DNA ladder 1 kb as positive control using 1.5 g agarose gel and was photographed using transmitted UV light and polaroid film. Results showed the PCR product indicated successful clones of the gDNA with gene primer sizes of 1500 bp to 1517 bp.

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