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Rapid Screening of Recombinant Plasmids by Direct Colony Quantitative Real-Time PCR
Department of Thyroid and Breast Surgery, General Hospital of Jinan Military Command, Jinan, China
Shandong Prosthetics Orthotics Rehabilitation Center, Jinan, China
Qilu Hospital of Shandong University, Jinan, China
Qilu Hospital of Shandong University, Jinan, China
Qianfo Mt. Hospital of Shandong University, Jinan, China
Qilu Hospital of Shandong University, Jinan, China
Liao City Municipal Hospital, Liaocheng, China
Department of Thyroid and Breast Surgery, General Hospital of Jinan Military Command, Jinan, China
- 1 Department of Thyroid and Breast Surgery, General Hospital of Jinan Military Command, Jinan, China
- 2 Shandong Prosthetics Orthotics Rehabilitation Center, Jinan, China
- 3 Qilu Hospital of Shandong University, Jinan, China
- 4 Qilu Hospital of Shandong University, Jinan, China
- 5 Qianfo Mt. Hospital of Shandong University, Jinan, China
- 6 Qilu Hospital of Shandong University, Jinan, China
- 7 Liao City Municipal Hospital, Liaocheng, China
- 8 Department of Thyroid and Breast Surgery, General Hospital of Jinan Military Command, Jinan, China
Advances in Bioscience and Biotechnology·Volume 07 (2016)·Pages 428–433·Published 29 September 2016·DOI10.4236/abb.2016.710041
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Abstract
Quantitative real-time PCR (qPCR) was applied to rapid screening of positive plasmid clones. Insert-specific primer pairs were used in qPCR colony screening, and false positive colonies could easily be distinguished from true positive ones by comparing their Ct values. In addition, qPCR is particularly suitable when amplicon is small (<150 bp). This method is sensitive, simple and fast, obviates the need for gel electrophoresis, and is a cost-effective alternative to the traditional PCR approach.
KeywordsqPCRColony PCRPlasmid Screening
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