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Dna Sequences Using Polymerase Chain Reaction

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Amplification of 16S Ribosomal DNA Sequences using Polymerase Chain Reaction
Edwina Abou Haidar,
Houssam Al Koussa,
Mary AbedAlAhad.
Department of Biology, Lebanese American University, Byblos, Lebanon

Abstract The 16s rRNA gene sequencing is a widely common amplicon sequencing method used to identify and compare bacteria in a given sample. This method is well established and allows to study phylogeny and taxonomy of complex microbiomes. In this study, an unknown sample of extracted microbial DNA was analyzed by performing the polymerase chain reaction followed by agarose gel electrophoresis. The results were accurate since three distinct bands (1500 basepairs) corresponded to our sample (duplicates) and the positive control. This indicates clearly the amplification of the 16s rRNA gene whose further sequencing technique constitutes a pivotal tool in the accurate identification of bacterial isolates as well as the discovery of novel bacteria in clinical microbiology laboratories.
Abbreviations
rDNA: ribosomal DNA rRNA: ribosomal RNA bp: base pair
PCR: Polymerase Chain Reaction Introduction Physiological and biochemical tests constituting the basis of conventional differentiation between bacterial species are somehow cumbersome, consume a lot of time and require different approaches [1]. Furthermore, the commercial identification systems failed to identify commonly encountered bacteria and uncommon isolates. In fact, these commercial systems

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