Indian Journal of Public Health Research & Development
Year : 2019, Volume : 10, Issue : 1
First page : ( 315) Last page : ( 320)
Print ISSN : 0976-0245. Online ISSN : 0976-5506.
Article DOI : 10.5958/0976-5506.2019.00062.7

Polymerase Chain Reaction (PCR) Method for Identification Gene Escherichia coli and Officer Depot Behavior in Drinking Water Refill

Baharuddin Alfina*

Departement of Environmental Health, University Moslem of Indonesia

*Corresponding Author: Alfina Baharuddin Departement of Environmental Health, University Moslem of Indonesia, Email: alfina.baharuddin@umi.ac.id

Online published on 21 February, 2019.

Abstract

Polymerase Chain Reaction (PCR) is one method for amplification (multiplication) of oligonucleotide primers directed enzymatically by specific DNA sequences. An accurate technique for molecular identification of bacteria is identification of the 16S-rRNA gene. Ribosomal RNA is most widely used as a molecular marker. The purpose of this study was to identify E. Coli bacterial 16S rRNA gene in refill drinking water. The research method used is analytic observational. As for the sample, there were 5 depots in Mariso sub-district and 5 depots in Panakkukang sub-district with a total sample of 3 samples measured on inlets, processes and outlets. The working principle of bacterial RNA is extracted and purified using RNX-plus kit. Deoxyribonuclease enzymes are used to remove DNA contamination from purified RNA. To remove the enzyme, 1 ml of tetra ethylene acetic acid (EDTA, 25 Mm) was added for 10 minutes at 65° C. Boom DNA extract method, DNA amplification by RT-PCR, PCR product detection, the results obtained in the form of RNA black band pattern (RNA band) where the results of electrophoresis were obtained by RNA bands (RNA bands) at 723 bp. The results showed that there was 1 consumer at the household level who was positive for E. coli bacteria. as a conclusion that the RNA genomic RT-PCR template can be used to detect Escherchia coli bacteria in refill drinking water more quickly, effectively and accurately when compared to culture methods which only require 4 hours of detection.

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Keywords

16S-RNA, RT-PCR, E.coli, 723 bp.

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