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http://krishi.icar.gov.in/jspui/handle/123456789/82107
Title: | Optimization of DNA isolation and PCR parameters in MyrislicI species and related genera for RAPD and ISSR analysis |
Other Titles: | Not Available |
Authors: | T E Sheeja George, Johnson K Jerome Jessy R Sandeep Varma S Syamkumar B Krishnamoorthy V A Parthasarathy |
ICAR Data Use Licennce: | http://krishi.icar.gov.in/PDF/ICAR_Data_Use_Licence.pdf |
Author's Affiliated institute: | ICAR::Indian Institute of Spices Research |
Published/ Complete Date: | 2008 |
Project Code: | Not Available |
Keywords: | Optimization DNA isolation PCR parameters Myristica sp RAPD ISSR analysis |
Publisher: | ICAR- Indian institute of spices research,Kozhikode, Kerala India |
Citation: | 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. |
Series/Report no.: | Not Available; |
Abstract/Description: | 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. |
Description: | Not Available |
ISSN: | Not Available |
Type(s) of content: | Research Paper |
Sponsors: | Not Available |
Language: | English |
Name of Journal: | Journal of Spices and Aromatic Crops |
Volume No.: | 17(2) |
Page Number: | 91-97 |
Name of the Division/Regional Station: | Not Available |
Source, DOI or any other URL: | Not Available |
URI: | http://krishi.icar.gov.in/jspui/handle/123456789/82107 |
Appears in Collections: | HS-IISR-Publication |
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