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Title: | Rapid and inexpensive NaOH based direct PCR for amplification of nuclear and organelle DNA from ramie (Boehmeria nivea), a bast fibre crop containing complex polysaccharides. |
Other Titles: | Not Available |
Authors: | P. Satya, S. Mitra, D.P. Ray, B.S. Mahapatra, M. Karan, S. Jana, A.K. Sharma |
ICAR Data Use Licennce: | http://krishi.icar.gov.in/PDF/ICAR_Data_Use_Licence.pdf |
Author's Affiliated institute: | ICAR::Central Research Institute for Jute and Allied Fibres |
Published/ Complete Date: | 2013-01-01 |
Project Code: | NFBSFARA FQ3030/2012-13 |
Keywords: | Ramie, Boehmeria nivea, Direct PCR, NaOH, Dilution, Polysaccharides |
Publisher: | Elsevier |
Citation: | Not Available |
Series/Report no.: | Not Available; |
Abstract/Description: | tMost of the PCR based approaches in plant science rely on lengthy and expensive DNA isolation protocols,which often involve use of hazardous chemicals. Direct PCR methods save time and cost of experi-ments and also increase efficiency of PCR. We have compared three rapid DNA extraction processes fordirect PCR in ramie (Boehmeria nivea), a fibre crop species containing high amount of gummy complexpolysaccharides and developed modified protocols for direct PCR using NaOH as extraction buffer andTris/Tris–HCl/Tris–EDTA as dilution buffer. These protocols were successful in amplification of nuclearDNA from leaf and stem tissues using ISSR and SSR markers and also chloroplast DNA amplification usingprimers for rbcL gene. Our results also show that nature and quantity of dilution buffer are importantfor increasing efficiency of direct PCR. The NaOH based methods are simpler, cheaper and economicalcompared to other direct PCR methods and work very well for tissues containing high amount of complexpolysaccharides. The protocols are suitable for batch processing and high throughput genotyping withina short time period, which will have many applications in plant genomic researches. |
Description: | Not Available |
ISSN: | Not Available |
Type(s) of content: | Journal |
Sponsors: | Indian Council of Agricultural Resaerch |
Language: | English |
Name of Journal: | Industrial Crops and Products |
Journal Type: | Research Journal |
NAAS Rating: | 10.24 |
Impact Factor: | 5.645 |
Volume No.: | 50 |
Page Number: | 532-536 |
Name of the Division/Regional Station: | Not Available |
Source, DOI or any other URL: | http://dx.doi.org/10.1016/j.indcrop.2013.07.049 |
URI: | http://krishi.icar.gov.in/jspui/handle/123456789/59114 |
Appears in Collections: | CS-CRIJAF-Publication |
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