Microsatellite markers from a microdissected swine chromosome 6 genomic library

Abstract

To develop additional microsatellite (MS) markers in the region of the porcine skeletal muscle ryanodine receptor gene (RYR1), a microdissected genomic library was generated from the proximal half of the q arm of swine chromosome 6. Purified DNA was restriction enzyme-digested, ligated to oligonucleotide adaptors and amplified by PCR using primers complementary to the adaptor sequences. The purity of the amplified products and boundaries of the microdissected chromosomal region were verified by fluorescence in situ hybridization. (CA)n-containing sequences were then identified in a small insert genomic library generated from the PCR-amplified microdissected DNA. Oligonucleotide primers were developed for the PCR amplification of 30 of the 46 (CA)n repeat-containing clones, which were subsequently used to amplify DNA isolated from unrelated pigs of different breeds to determine the informativeness of these MS markers. Twenty-two of these MS markers were genotyped on the University of Illinois Yorkshire x Meishan swine reference population. These 22 markers were all assigned within a 50.7-CM region of the swine chromosome 6 linkage map, indicating the specificity of the microdissected library.

Variable Numbers of TTC Repeats inMycobacterium lepraeDNA from Leprosy Patients and Use in Strain Differentiation

“…Likewise, three nucleotide repeats, CAA/CAG, have been employed in the typing of Candida albicans strains (2). In addition, such short DNA repeats, also known as microsatellites, have been widely used in forensic investigation (15) and for determining the genetic traits of domestic and pet animals, including horses (1), pigs (26), chickens (14), and dogs (9).…”

Development and mapping of microsatellites from a microdissected BTA 11‐specific DNA library

“…1997; Sonstegard et al. 1997; Zhao et al . 1999) have described the successful use of chromosome‐specific libraries for developing microsatellite markers to increase marker density on specific chromosomes.…”

High-resolution genetic map of bovine chromosome 29 through focused marker development

“…DNA libraries generated by chromosome flow sorting or microdissection provide a rich source of markers for specific chromosomes. Zhao et al (1999) recently reported 22 new markers from a microdissected library of porcine chr 6 (SSC6) and Wang et al (2000) isolated 31 markers from an SSC8 library. Recently, Ambady et al (2001) isolated 17 new ms markers from a BTA11-specific library and Liu et al (in press) obtained 34 new ms markers from a BTAY library.…”