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PCR-suppression effect: kinetic analysis and application to representative or long-molecule biased PCR-based amplification of complex samples.

Authors
  • Dai, Zhong-Min
  • Zhu, Xiao-Jing
  • Chen, Qi
  • Yang, Wei-Jun
Type
Published Article
Journal
Journal of Biotechnology
Publisher
Elsevier
Publication Date
Feb 20, 2007
Volume
128
Issue
3
Pages
435–443
Identifiers
PMID: 17194496
Source
Medline
License
Unknown

Abstract

In the present study, we analyzed the kinetics of the PCR-suppression effect (PS-effect) and firstly demonstrate that lower annealing temperature enhances PS-effect. Furthermore, we controlled the average size of complex PCR products over a wide range, and simultaneously amplified targets from 0.25 to 10kb by regulating the degree of suppression in a single-primer PCR. In addition, we describe an improved template-switching full-length cDNA synthesis method that greatly reduces truncated cDNA. This study provides a general guide for the design of PS-effect related PCR and is useful for representative or long-transcript enriched cDNA library construction, especially when only a small amount of total RNA is available.

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