ICAR Research Data Repository for Knowledge Management
Evaluation of PCR-based methods for isolating flanking regions of genes.
IR@CSIR-CFTRI
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http://ir.cftri.com/1947/
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| Title |
Evaluation of PCR-based methods for isolating flanking regions of genes.
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| Creator |
Satyanarayana, K. V.
Chandrashekar, A. Ravishankar, G. A. |
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| Subject |
03 Biochemistry & Molecular Biology
05 Tissue Culture 02 Evolution & genetics |
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| Description |
Several polymerase chain reaction (PCR)-based methods are available for isolation of unknown genomic fragments. In the present study, a comparative evaluation of a few methods of ligation-mediated PCR methods and a ligation-independent one were made by isolating promoter fragment for N-methyltransferase gene involved in the caffeine biosynthetic pathway of Coffea canephora. The benefits of tertiary PCR and the effects of a 4-base cutting restriction endonuclease on the size of the PCR products obtained were demonstrated in one of the ligation-mediated PCR methods. The methods adopted in this study differed in the sizes of the 5'-flanking regions obtained. The efficiencies of various methods used reflect the inherent limitations of the PCR-based methods for isolation of unknown flanking regions.
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| Date |
2006
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| Type |
Article
NonPeerReviewed |
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| Format |
application/pdf
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| Language |
en
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| Rights |
—
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| Identifier |
http://ir.cftri.com/1947/1/MolBiotech32111-116.pdf
Satyanarayana, K. V. and Chandrashekar, A. and Ravishankar, G. A. (2006) Evaluation of PCR-based methods for isolating flanking regions of genes. Molecular Biotechnology, 32. pp. 111-116. ISSN 1073-6085 |
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