title: SiteFinding-PCR: a simple and efficient PCR method for chromosome walking.
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.
authors.affiliation: The National Laboratory of Protein Engineering and Plant Genetic Engineering, College of Life Sciences, Peking University Beijing 100871, China.
authors.forename: Guihong
authors.initials: G
authors.lastname: Tan
authors.forename: Yin
authors.initials: Y
authors.lastname: Gao
authors.forename: Miao
authors.initials: M
authors.lastname: Shi
authors.forename: Xinyue
authors.initials: X
authors.lastname: Zhang
authors.forename: Shanping
authors.initials: S
authors.lastname: He
authors.forename: Zhangliang
authors.initials: Z
authors.lastname: Chen
authors.forename: Chengcai
authors.initials: C
authors.lastname: An
mesh_terms: D017360:Arabidopsis / Q000235:genetics; D016386:Chromosome Walking / Q000379:methods*; D000458:Cyanobacteria / Q000821:virology; D017931:DNA Primers / Q000737:chemistry; D004269:DNA, Bacterial / Q000235:genetics; D016254:Mutagenesis, Insertional; D017900:Myoviridae / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D012231:Rhizobium / Q000235:genetics
chemical_list: D017931:DNA Primers; D004269:DNA, Bacterial