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

Paper ID Volume ID Publish Year Pages File Format Full-Text
25264 43564 2007 9 PDF Available
Title
PCR-suppression effect: Kinetic analysis and application to representative or long-molecule biased PCR-based amplification of complex samples
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 10 kb 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.

Keywords
PCR-suppression effect; Annealing temperature; Complex PCR; Representation; Full-length cDNA
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PCR-suppression effect: Kinetic analysis and application to representative or long-molecule biased PCR-based amplification of complex samples
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 128, Issue 3, 20 February 2007, Pages 435–443
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us