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Optimization of DNA isolation and PCR parameters in MyrislicI species and related genera for RAPD and ISSR analysis

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Title Optimization of DNA isolation and PCR parameters in MyrislicI species and related genera for RAPD and ISSR analysis
Not Available
 
Creator T E Sheeja
George, Johnson K
Jerome Jessy
R Sandeep Varma
S Syamkumar
B Krishnamoorthy
V A Parthasarathy
 
Subject Optimization
DNA isolation
PCR parameters
Myristica sp
RAPD
ISSR analysis
 
Description Not Available
An efficient protocol for isolation of DNA from wild and related genera of Myristica rich inpolysaccharides and polyphenols was developed. The protocol utilizes CTAB (3%), 1.5% PVPand 0.3% s-mercaptoethanol for isolation and RNase and phenol chloroform extraction forpurification. The yield of DNA ranged from 25-175 µg/g of fresh leaf tissue, with Knemaandamanica giving the highest yield. The present method yielded 10 times higher than theold methods. Characteristic patterns were generated on digestion of DNA by EcorI and HindIII restriction enzymes. PCR parameters were optimized using random primers (OPERONTechnology, USA). DNA concentration at 20 ng/reaction, annealing temperature of 45°C, 0.3mM dNTP in presence of 0.5 U of Taq DNA polymerase, and 2.0 mM MgCl2 and MJ ResearchGene Thermocycler was best. Successful amplification by ISSR and RAPD primers indicatedthat DNA is of good quality and free of polysaccharides and polyphenols.
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Date 2024-04-17T14:14:28Z
2024-04-17T14:14:28Z
2008
 
Type Research Paper
 
Identifier Sheeja, T.E., Johnson George, K., Jessy Jerome, Sandeep Varma, R., Syamkumar, S., Krishnamoorthy. B. and Parthasarathy, VA. (2008) Optimization of DNA isolation and PCR parameters in MyrislicI species and related genera for RAPD and ISSR analysis. Journal (~l Spices and Aromatic Crops) 17(2):9 1-97.
Not Available
http://krishi.icar.gov.in/jspui/handle/123456789/82107
 
Language English
 
Relation Not Available;
 
Publisher ICAR- Indian institute of spices research,Kozhikode, Kerala India