PCR hot-start using duplex primers
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Polymerase Chain Reaction: Basic Protocol Plus Troubleshooting and Optimization Strategies
Published on: May 22, 2012
Deming Kong1, Hanxi Shen, Yanping Huang
1State Key Laboratory, of Functional Polymer Materials for Adsorption and Separation, Chemical School, Nankai University, Tianjin 300071, P.R. China. kongdem@eyou.com
Biotechnology Letters
|April 2, 2004
Abstract:
A new technique of PCR hot-start using duplex primers has been developed which can decrease the undesirable products arising throughout PCR amplification thereby giving better results than a manual hot-start method.
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Rapid Amplification of cDNA Ends, or RACE, is one of the most effective methods to obtain a full-length cDNA from an mRNA sequence between a known internal region to the unknown sequence at the 5’ or 3’ end. The unknown region is cloned in the cDNA by a gene-specific primer that binds the known end, and a hybrid primer that attaches a predefined anchor sequence to the unknown end of the cDNA. The sequence in between is amplified by PCR with an anchor primer and a gene-specific primer.
Since the...
Since the...