Lab Report

Authors: Nilantha Gamalathge

Abstract Bifidobacteria Identification is important in medical and pharmaceutical industry. The molecular analytic technique such as PCR gel electrophoresis and MALDI –TOF-MS are high sensitive and useful technique in identifying Bifidobacteria. This practical was planned to identification of Bifidobacteria by both method. PCR method is based on denaturing, annealing and extension of target DNA in the bacteria in vitro with other requirements like taq polymerase enzyme, DNA primers, Mg etc with temperature control. MALDI –TOF-MS is based on analyet Matrix co crystallization and firing of laser beam get ionization and detection time of flight by mass spectrometry Un known 3 cutler plats (S1, S2,S3) were used for both methods and correctly identified up to genus level by PCR agarose gel electrophoresis. MALDI TOF MS was done in direct application and formic acid extraction method. Both ways identified up to bifid bacteria species and lactobacillus. Both methods were equally highly sensitive to identify Bifidobacteria. Identification of species was highly accurate MALDY TOF –MS method. But both methods have done with limited sample size. Both methods are complex, high cost and need high technical knowledge. MALDI method results depend on software apart from other steps. PCR can improve to identify Bifid bacteria up to species level with the use of species-level primers.MALDI method with formic acid exaction can improve sensitivity with the purification of the protein. (PCR-polymerase Chain Reaction.MALDI-TOF-MS- Matrix-Assisted Laser Desorption Ionization Time of Flight Mass Spectrometry).

Comments: 19 Pages.

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[v1] 2018-08-04 02:48:56

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