A genomic walking method for screening sequence length polymorphism

作者：Walser Jean Claude^*; Evgen'ev Michael B; Feder Martin E

来源：Molecular Ecology Notes, 2006, 6(2): 563-567.

DOI：10.1111/j.1471-82826.2006.02176.x

• heat-shock genes
• high-throughput screening
• insertion
• promoter regulation
• transposable elements
• universal fast walking

We adapted a recently developed nonrestrictional, nonligational genome walking method, Universal Fast Walking (UFW), for detection of length polymorphism in the proximal promoter region of genes. We demonstrate its efficacy at discovering naturally occurring transposition into heat-shock genes of wild Drosophila and show that it surmounts limitations of simple polymerase chain reaction (PCR) approaches. We further present modifications to the standard UFW protocol and provide some guidelines to improve specificity. Although the resultant banding pattern of a standard UFW can be regarded as a DNA fingerprint, many amplicons result from false priming and not real polymorphisms. We describe ways to distinguish between UFW amplicons and false priming products in a high-throughput assay.

• 出版日期2006-6