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

@article{citeulike:1025092, 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.}, author = {Dai, Zhong-Min and Zhu, Xiao-Jing and Chen, Qi and Yang, Wei-Jun}, citeulike-article-id = {1025092}, doi = {10.1016/j.jbiotec.2006.10.018}, journal = {Journal of Biotechnology}, keywords = {cdna, cloning, construction, library}, posted-at = {2007-01-04 15:36:34}, priority = {2}, title = {PCR-suppression effect: Kinetic analysis and application to representative or long-molecule biased PCR-based amplification of complex samples}, url = {http://dx.doi.org/10.1016/j.jbiotec.2006.10.018}, volume = {In Press, Corrected Proof} }

TY - JOUR ID - citeulike:1025092 L3 - citeulike-article-id:1025092 N2 - 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. JF - Journal of Biotechnology TI - PCR-suppression effect: Kinetic analysis and application to representative or long-molecule biased PCR-based amplification of complex samples VL - In Press, Corrected Proof KW - cdna KW - cloning KW - construction KW - library AU - Dai, Zhong-Min AU - Zhu, Xiao-Jing AU - Chen, Qi AU - Yang, Wei-Jun UR - http://dx.doi.org/10.1016/j.jbiotec.2006.10.018 ER -