NISCAIR Online Periodicals Repository

NISCAIR ONLINE PERIODICALS REPOSITORY (NOPR)  >
NISCAIR PUBLICATIONS >
Research Journals >
Indian Journal of Biotechnology (IJBT) >
IJBT Vol.09 [2010] >
IJBT Vol.09(1) [January 2010] >


Title: Evaluation and optimization of DNA extraction method for Dalbergia sissoo leaf
Authors: Ginwal, H S
Maurya, Shalini Singh
Keywords: CTAB
Dalbergia sissoo
DNA extraction
forest tree
RAPD
ISSR
Issue Date: Jan-2010
Publisher: CSIR
Abstract: A modified protocol for Dalbergia sissoo genomic DNA isolation has been optimized based on a cetyl trimethyl ammonium bromide (CTAB) method, described for other forest species. Leaves obtained from macro-propagated clones and mature trees of D. sissoo were tested. The method involves mortar grinding of tissue, a modified CTAB extraction buffer incorporating high salt concentrations, polyvinyl pyrrolidone and successive isoamyl alcohol-chloroform extractions with modified temperature conditions. The modification involved the use of doubled concentration of polyvinyl pyrrolidone (4% instead of 2%), increased incubation time with extraction buffer (40 min instead of 30 min), use of freshly prepared CTAB buffer and increased timing of washing of DNA pellet with wash buffer (45 min instead of 30 min). The yield was approximately 100 to 400 µg DNA per 100 mg of leaf tissue. The genomic DNA obtained by this method was suitable to be used in RAPD and ISSR analysis. This extraction method would allow the molecular analysis of DNA from different clones of D. sissoo.
Page(s): 69-73
ISSN: 0975-0967 (Online); 0972-5849 (Print)
Source:IJBT Vol.09(1) [January 2010]

Files in This Item:

File Description SizeFormat
IJBT 9(1) 69-73.pdf500.91 kBAdobe PDFView/Open
 Current Page Visits: 1377 
Recommend this item

 

National Knowledge Resources Consortium |  NISCAIR Website |  Contact us |  Feedback

Disclaimer: NISCAIR assumes no responsibility for the statements and opinions advanced by contributors. The editorial staff in its work of examining papers received for publication is helped, in an honorary capacity, by many distinguished engineers and scientists.

CC License Except where otherwise noted, the Articles on this site are licensed under Creative Commons License: CC Attribution-Noncommercial-No Derivative Works 2.5 India

Copyright © 2012 The Council of Scientific and Industrial Research, New Delhi. All rights reserved.

Powered by DSpace Copyright © 2002-2007 MIT and Hewlett-Packard | Compliant to OAI-PMH V 2.0

Home Page Total Visits: 585405 since 06-Feb-2009  Last updated on 17-Sep-2014Webmaster: nopr@niscair.res.in