<em>Bst</em> DNA Polymerase Large Fragment for Whole Genome Amplification

doi:10.13523/j.cb.20160708

China Biotechnology

2016, Vol. 36

Issue (7): 48-54 DOI: 10.13523/j.cb.20160708

Bst DNA Polymerase Large Fragment for Whole Genome Amplification

ZHANG Xin-yuan^1,2, DENG Jia^1,2, LIU Xin^1,2, CHEN Lin^1,2, LIN Jun^1,2, CAI Wei-wen^1,2

1 Institute of Applied Genomics, Fuzhou University, Fuzhou 350108, China;
2 College of Biological Science and Engineering, Fuzhou University, Fuzhou 350108, China

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Abstract

As a common DNA polymerase, Bst DNA polymerase large fragment has been a kind of important DNA multiple displacement amplification enzyme, which can cause chain reaction, high fidelity, high temperature resistance and so on. Objective:Formalin-fixed paraffin-embedded (FFPE) tissues represent the largest source of archival biological material available for genomic studies of human cancer. Therefore, it is desirable to develop methods that enable whole genome amplification (WGA) using DNA extracted from FFPE tissues. In order to reduce cost, a high yield, convenient and high activity of Bst DNA polymerase large fragment expression system had been designed, and the conditions in the amplification of the gastric cancer genomic DNA from FFPE tissues were explored. Methods:Bst DNA polymerase large fragment was cloned and expressed by the PTWIN1 plasmid, and purified by the chintin column. It was used to amplify the Human genome DNA at different temperature to explore the optimum temperature. Then the gastric cancer genomic DNA from FFPE tissues was amplified on the optimum condition. Results:The Bst DNA polymerase large fragment yielded a median 200-fold of DNA amplification. Conclusion:Bst DNA polymerase large fragment is suitable for WGA of DNA extracted from FFPE tissues. Amplified DNA may be used for the detection of gene copy number changes and genome profiling by array CGH.

Key words： Multiple displacement amplification Bst DNA polymerase large fragment Formalin-fixed Whole genome amplification paraffin-embedded

Received: 28 December 2015 Published: 02 March 2016

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	ZHANG Xin-Yuan
	DENG Jia
	LIU Xin
	CHEN Lin
	LIN Jun
	SA Wei-Wen

Cite this article:

ZHANG Xin-yuan, DENG Jia, LIU Xin, CHEN Lin, LIN Jun, CAI Wei-wen. Bst DNA Polymerase Large Fragment for Whole Genome Amplification. China Biotechnology, 2016, 36(7): 48-54.

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https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20160708 OR https://manu60.magtech.com.cn/biotech/Y2016/V36/I7/48

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