Optimized adaptor polymerase chain reaction method for efficient genomic walking.

Abstract:

:Genomic walking is one of the most useful approaches in genome-related research. Three kinds of PCR-based methods are available for this purpose. However, none of them has been generally applied because they are either insensitive or inefficient. Here we present an efficient PCR protocol, an optimized adaptor PCR method for genomic walking. Using a combination of a touchdown PCR program and a special adaptor, the optimized adaptor PCR protocol achieves high sensitivity with low background noise. By applying this protocol, the insertion sites of a gene trap mouse line and two gene promoters from the incompletely sequenced Xenopus laevis genome were successfully identified with high efficiency. The general application of this protocol in genomic walking was promising.

authors

Xu P,Hu RY,Ding XY

doi

10.1111/j.1745-7270.2006.00194.x

subject

Has Abstract

pub_date

2006-08-01 00:00:00

pages

571-6

issue

8

eissn

1672-9145

issn

1745-7270

journal_volume

38

pub_type

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