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Optimization of DNA isolation and peR parameters in Myristica sp. and related gen­ era for RAPD and ISSR analysis.

DSpice at Indian Institute of Spices Research

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Field Value
 
Creator SHEEJA, T E
JOHNSON, GEORGE K
JESSY JEROME
SANDEEP VARMA, R
SYAMKUMAR, S
KRISHNAMOORTHY, B
PARTHASARATHY, V A
 
Date 2010-08-09T06:28:50Z
2010-08-09T06:28:50Z
2008
 
Identifier Journal of Spices and Aromatic Crops Volume 17 (2) : 91-97 (2008)
http://hdl.handle.net/123456789/532
 
Description An efficient protocol for isolation of DNA from wild and related genera of Myristica rich in polysaccharides and polyphenols was developed. The protocol utilizes CTAB (3%), 1.5% PVP and 0.3% B-mercaptoethanol for isolation and RNase and phenol chloroform extraction for purification. The yield of DNA ranged from 25-175 flg/g of fresh leaf tissue, with Knema andamanica giving the highest yield. The present method yielded 10 times higher than the old methods. Characteristic patterns were generated on digestion of DNA by Ecod and Hind III restriction enzymes. PCR parameters were optimized using random primers (OPERON Technology, USA). DNA concentration at 20 ng/reaction, annealing temperature of 45°C, 0.3 mM dNTP in presence of 0.5 U of Taq DNA polymerase, and 2.0 mM MgCl and MJ Research z Gene Thermocycler was best. Successful amplification by ISSR and RAPD primers indicated that DNA is of good quality and free of polysaccharides and polyphenols.
 
Format 491594 bytes
application/pdf
 
Language en
 
Publisher Indian Society for Spices
 
Subject DNA isolation
optimization
PCR
polymorphism
restriction digestion
 
Title Optimization of DNA isolation and peR parameters in Myristica sp. and related gen­ era for RAPD and ISSR analysis.
 
Type Article