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SiteFinding-PCR: a simple and efficient PCR method for chromosome walking.

Guihong Tan·Yin Gao·Miao Shi·Xinyue Zhang·Shanping He·Zhangliang Chen·Chengcai An
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Abstract

In this paper, we present a novel PCR method, termed SiteFinding-PCR, for gene or chromosome walking. The PCR was primed by a SiteFinder at a low temperature, and then the target molecules were amplified exponentially with gene-specific and SiteFinder primers, and screened out by another gene-specific primer and a vector primer. However, non-target molecules could not be amplified exponentially owing to the suppression effect of stem-loop structure and could not be screened out. This simple method proved to be efficient, reliable, inexpensive and time-saving, and may be suitable for the molecules for which gene-specific primers are available. More importantly, large DNA fragments can be obtained easily using this method. To demonstrate the feasibility and efficiency of SiteFinding-PCR, we employed this method to do chromosome walking and obtained 16 positive results from 17 samples.

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ArabidopsisChromosome WalkingCyanobacteriaDNA PrimersDNA, BacterialMutagenesis, InsertionalMyoviridaePolymerase Chain ReactionRhizobiumEvaluation StudyJournal ArticleResearch Support, Non-U.S. Gov't

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Guihong Tan, Yin Gao, Miao Shi, Xinyue Zhang, Shanping He, Zhangliang Chen, Chengcai An. 2005-08-02. SiteFinding-PCR: a simple and efficient PCR method for chromosome walking.. https://doi.org/10.1093/nar%2Fgni124

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