Method for Improving Rolling Circle Amplification Sensitivity and Specificity
Science Discovery
Volume 5, Issue 6, November 2017, Pages: 496-501
Received: Dec. 27, 2017; Published: Dec. 28, 2017
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Authors
Yun Wei Zhang, School of Biomedicial Sciences, Huaqiao University, Quanzhou, China
Dan Hou, Key Laboratory of Oral Medicine, Guangzhou Institute of Oral Disease, Stomatology Hospital of Guangzhou Medical University, Guangzhou, China
Wei Dong Xiao, School of Biomedicial Sciences, Huaqiao University, Quanzhou, China
Jenni Firrman, Dairy and Functional Foods Research Units, Eastern Regional Research Center, Agriculture Research Service, United States Department of Agriculture, Wyndmoor, The United States of America
Lin Shu Liu, Dairy and Functional Foods Research Units, Eastern Regional Research Center, Agriculture Research Service, United States Department of Agriculture, Wyndmoor, The United States of America
Yong Diao, School of Biomedicial Sciences, Huaqiao University, Quanzhou, China
Zhi Qing Qi, School of Biomedicial Sciences, Huaqiao University, Quanzhou, China
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Abstract
Rolling circle amplification (RCA) of genomic or plasmid DNA has found wide used in technique academic and biotechnology reseach. In this study, we have successfully found what caused the nonspecific product in the reaction, thus designing an improved RCA which overcomes false priming at sites where primer dimers are formed. The results demonstrated that PEG or PEI could interact with the components of the improved RCA system, and help increase the reaction rate and yield of the improved RCA system, and dramatically reduces the nonspecific RCA products. This low-cost and efficient method dramatically overcomes nonspecific product amplification phenomenon in the RCA reaction which is problematic and has existed for a long time.
Keywords
Rolling Circle Amplification, Recombinant Plasmid, Polyetherimide, Polyethylene Glycol
To cite this article
Yun Wei Zhang, Dan Hou, Wei Dong Xiao, Jenni Firrman, Lin Shu Liu, Yong Diao, Zhi Qing Qi, Method for Improving Rolling Circle Amplification Sensitivity and Specificity, Science Discovery. Vol. 5, No. 6, 2017, pp. 496-501. doi: 10.11648/j.sd.20170506.28
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