Rapid and efficient generation of pcr templates from Escherichia coli, Saccharomyces cerevisiae and Oryza sativa using a microwave and by boiling

被引:4
作者
He, R
Wang, Q
Zhang, Y
Liu, QY [1 ]
Li, G
Cui, L
Wang, ZX
Cao, Y
Li, ZJ
Cheng, D
Zhang, WQ
Li, BJ
Pang, Y
机构
[1] Zhongshan Univ, Key Lab Gene Engn Minist Educ, Guangzhou 510275, Peoples R China
[2] Zhongshan Univ, Biotechnol Res Ctr, Guangzhou 510275, Peoples R China
[3] Zhongshan Univ, Natl Key Lab Biocontrol, Guangzhou 510275, Peoples R China
[4] Beijing Union Med Coll Hosp, Inst Med Biotechnol, Beijing 100730, Peoples R China
关键词
D O I
10.1111/j.1745-4581.2005.00003.x
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Simple methods for the preparation of polymerase chain reaction (PCR) templates from Escherichia coli, Saccharomyces cerevisiae and Oryza sativa using a microwave and by boiling are described without the need for any special chemical reagent such as a detergent. The microwave and boiling methods were robust for the preparation of both plasmid and genomic templates of E. coli, as was the boiling approach for S. cerevisiae DNA. The microwave procedure was only suitable for dry yeast colonies with fair efficiency as absence of water (H2O) was critical for a successful PCR. Oryza sativa genomic DNA was amplified with both approaches with satisfactory efficiency Escherichia coli plasmids can be prepared using a microwave with a yield equivalent to 15-20% of an alkaline lysis mini-prep. PCR templates were also efficiently generated from Pichia pastoris and Ganoderma lucidum by using the boiling method.
引用
收藏
页码:19 / 28
页数:10
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