Record Details

Novel primer designing and PCR-AFLP approach for an expeditious detection of coliforms in potable waters

NOPR - NISCAIR Online Periodicals Repository

View Archive Info
 
 
Field Value
 
Title Novel primer designing and PCR-AFLP approach for an expeditious detection of coliforms in potable waters
 
Creator Shanker, A Shiva
Pindi, Pavan Kumar
 
Subject 16S rRNA
Coliform
Molecular marker
Primer design
Restriction enzymes
 
Description 65-75
Molecular techniques involving 16S rRNA gene have long been proved to be a mainstay of sequence-based bacterial analysis and enhance the competence of bacterial removal in drinking water and food. The main goal of this analysis was to reduce the time of detection of total coliforms by developing 16S rRNA based DNA markers by targeting variable region in the 16S rRNA gene position of V2 and V9. Coliform specific primers (189F and 1447R) were designed to amplify total coliform with an amplicon size of 1300 bp. The PCR product was later digested with Hind III and BseRI (restriction enzymes) to differentiate the type of contamination caused by fecal and non-fecal coliforms respectively. The digested amplicons were run on agarose gel electrophoresis and contamination levels were estimated based on the respective band pattern. This method can be applicable to know the coliform contamination levels of potable waters, in food and beverage industries within a short period of time. To our knowledge, this is the first report on newly designed primers which not only amplify coliform bacteria, followed by various restriction digestions of these amplicons but also provides unique band patterns to identify coliforms at genus level.
 
Date 2023-01-10T09:17:02Z
2023-01-10T09:17:02Z
2023-01
 
Type Article
 
Identifier 0975-0959 (Online); 0301-1208 (Print)
http://nopr.niscpr.res.in/handle/123456789/61183
https://doi.org/10.56042/ijbb.v60i1.69576
 
Language en
 
Publisher NIScPR-CSIR, India
 
Source IJBB Vol.60(01) [January 2023]