1. Method for Improving Rolling Circle Amplification Sensitivity and Specificity
- Author
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Lin Shu Liu, Yun Wei Zhang, Zhi Qing Qi, Yong Diao, Dan Hou, Jenni Firrman, and Weidong Xiao
- Subjects
Plasmid dna ,Rolling circle replication ,Chemistry ,Primer dimer ,Biophysics ,General Medicine ,Sensitivity (control systems) - 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.
- Published
- 2017