Aug 05, 2020

Public workspaceDNA extraction and Sequencing

DOI
dx.doi.org/10.17504/protocols.io.bjdxki7n
  • 1domnic colvin
  • UM-DAE Centre for Excellence in Basic Sciences
avinash.kale
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DOI: dx.doi.org/10.17504/protocols.io.bjdxki7n
Document Citation: avinash.kale 2020. DNA extraction and Sequencing. protocols.io https://dx.doi.org/10.17504/protocols.io.bjdxki7n
License: This is an open access document distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Created: August 05, 2020
Last Modified: August 05, 2020
Document Integer ID: 40087
DNA extraction and Sequencing
The four bacteria which showed differential toxicity against the Culex quinquefasciatus were subjected to DNA isolation and purification through HiPurA™ Bacterial Genomic DNA Purification Spin Kit (Himedia). Identification of species was done by using 16S rRNA sequencing at Eurofins Genomics India Pvt. Ltd using three primers 27F, 1492R and 519R and further for confirmation at National Centre for Microbial Resources, (NCMR) - National Centre for Cell Science (NCCS- Pune, India) using five primers 27F, 1492R(18)(19), 704F, 907R and 519R(20)(21). The sequencing results obtained were analyzed and identified using the NCBI 16S rRNA bacterial database (https://blast.ncbi.nlm.nih.gov/Blast).