Submit Manuscript

Asian Journal of Animal Sciences

Year: 2011 | Volume: 5 | Issue: 3 | Page No.: 166-174
Facebook Twitter Digg Reddit Linkedin StumbleUpon E-mail

Article Trend

Total views 212

Authors

P. Prashant Chaudhary

S. Kumar Sirohi

S. Kumar

Keywords

Research Article

Improved Extraction of Quality DNA from Methanogenic Archaea Present in Rumen Liquor for PCR Application

P. Prashant Chaudhary, S. Kumar Sirohi and S. Kumar
The present study has been planned to standardize a simple and effective method for the isolation of good quality as well as quantity of methanogenic DNA in total genomic DNA from rumen liquor of Bubalus bubalis. Methanogens are a diverse group of organisms found in anaerobic environments such as anaerobic sludge digester, wet wood of trees, sewage, rumen, black mud, black sea sediments , etc which utilize carbon dioxide and hydrogen and produce methane. Methanogens exhibit great diversity in cell envelopes, ranging from simple, non rigid surface layers consisting of protein or glycoprotein subunits to a rigid “pseudomurein” sacculus, analogous to eubacterial murein. Methanogens having different chemical composition of cell wall and known for tough cell wall which is difficult break to isolate good quality and quantity of genomic DNA. Various DNA extraction methodologies have been used but problems are most often encountered in terms of low DNA yields and quality of DNA for further application. Method of DNA isolation based on guanidine thiocynate lysis buffer was compared with commonly used phenol chloroform method and commercial kit based methods, results showed that modified protocol generated high molecular weight genomic DNA while the other two methods resulted in considerable DNA degradation. Further, the isolated genomic DNA was tested for downstream applications such as PCR and Real-Time PCR using methanogens specific primers. In both the cases, the genomic DNA isolated by our protocol was comparatively better than rest of two protocols tested.
PDF Fulltext XML References Citation

Leave a Comment

Your email address will not be published. Required fields are marked *