Davis M Wayne, Hammarlund Marc, Harrach Tracey, Hullett Patrick, Olsen Shawn, Jorgensen Erik M
Department of Biology, University of Utah, Salt Lake City, Utah 84112-0840, USA.
BMC Genomics. 2005 Sep 12;6:118. doi: 10.1186/1471-2164-6-118.
In C. elegans, single nucleotide polymorphisms (SNPs) can function as silent genetic markers, with applications ranging from classical two- and three-factor mapping to measuring recombination across whole chromosomes.
Here, we describe a set of 48 primer pairs that flank SNPs evenly spaced across the C. elegans genome and that work under identical PCR conditions. Each SNP in this set alters a DraI site, enabling rapid and parallel scoring. We describe a procedure using these reagents to quickly and reliably map mutations. We show that these techniques correctly map a known gene, dpy-5. We then use these techniques to map mutations in an uncharacterized strain, and show that its behavioral phenotype can be simultaneously mapped to three loci.
Together, the reagents and methods described represent a significant advance in the accurate, rapid and inexpensive mapping of genes in C. elegans.
在秀丽隐杆线虫中，单核苷酸多态性（SNP）可作为沉默遗传标记，其应用范围从经典的双因子和三因子作图到测量整条染色体的重组。
在此，我们描述了一组48对引物，这些引物位于秀丽隐杆线虫基因组中均匀间隔的SNP两侧，并且在相同的PCR条件下有效。该组中的每个SNP都会改变一个DraI位点，从而实现快速并行评分。我们描述了一种使用这些试剂快速可靠地定位突变的方法。我们表明这些技术正确地定位了一个已知基因dpy - 5。然后我们使用这些技术定位一个未表征菌株中的突变，并表明其行为表型可同时定位到三个位点。
总之，所描述的试剂和方法代表了秀丽隐杆线虫基因精确、快速且廉价定位方面的重大进展。
