|Hawkins, Greg - EPICENTER, MADISON, WI|
|Bishop, Michael - ABS GLOBAL, DEFOREST, WI|
Submitted to: Animal Genetics
Publication Type: Abstract Only
Publication Acceptance Date: January 11, 1997
Publication Date: N/A
Technical Abstract: Source: A bovine genomic library in SuperCos 1 (Stratagene, La Jolla, Calif.) was screened with an end-labeled (GT)8 oligonucleotide probe according to Hawkins et al. (1995). PCR primers: Primer sequences and annealing temperatures are presented in Table 1. PCR conditions: PCR conditions contained 100 ng DNA, 15 mM MgCl2, 50 mM KCL, 10 mM Tris-HCL (pH8.3), 30 (mu)M dCTP, dGTP and dTTP, 3 (mu)M dATP, .1 (mu)Ci [(alpha)32P] dATP (3000 Ci/(mu)mole), .4 (mu)M for each of two primers, and .35 unit Taq polymerase. The PCR protocol was: initial denaturation, 94 degrees for 3 min followed by 30 cycles of 1 min at 94 degrees, 30 sec at the annealing temperature and 1 min at 72 degrees. The final extension at 72 degrees was for 4 min. Alleles: PCR products were analyzed and scored according to Bishop et al. (1994). The number and size of the alleles observed in parents (n=29) of the USDA/MARC reference families are summarized in Table 1. All eleven markers followed Mendelian inheritance patterns. An unamplified allele was observed for marker BMC4228. A relative score indicating the ease of genotyping is also presented in Table 1. Chromosomal location: Determined by linkage analysis (Table 1).