DNA NICKING FAVORS PCR RECOMBINATION

被引:36
作者
MARTON, A [1 ]
DELBECCHI, L [1 ]
BOURGAUX, P [1 ]
机构
[1] UNIV SHERBROOKE,FAC MED,DEPT MICROBIOL,SHERBROOKE J1H 5N4,QUEBEC,CANADA
基金
英国医学研究理事会;
关键词
D O I
10.1093/nar/19.9.2423
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
We attempted to use the polymerase chain reaction (PCR) to monitor in vitro recombination in a plasmid containing directly repeated sequences. Some of the plasmid preparations which had not been exposed to recombination conditions were however found to behave in the PCR test as if they had undergone homologous recombination. We show here that such false positives are attributable to a small degree of nicking and/or breaking of the DNA template. Presumably, such damage allows the formation of hybrid parental duplexes containing at least one truncated strand, the 3' end of which maps within the homology; extension of this 3' end by the polymerase then results in a linkage of sequences identical to that arising from homologous recombination.
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页码:2423 / 2426
页数:4
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