Rat Anti-Human GM-CSF Heterohybridoma [CWE3-32D22.4]

Allele and genotype frequencies for 3 tetrameric short tandem repeat loci were determined in a Swiss population sample (n = 100) using the GenePrint STR Multiplex System, electrophoresis of the PCR products in DNA sequencing gels and subsequent detection of allelic fragments by silver staining. The loci are HUMTHO1, TPOX, and CSF1PO. The observed heterozygosities are 83.0%, 60.0%, and 72.0%, respectively. The discrimination power determined for the individual loci is 0.914, 0.780, and 0.860, respectively, and the combined discrimination power for the tripler is 0.997. All loci meet Hardy Weinberg expectations and after Bonferroni correction there was no evidence that the population sample deviates from expectations of independence. Moreover, independence of alleles at these STR loci with other PCR-based loci derived from the same Swiss population sample, previously reported, were considered. These loci were DQA1, LDLR, GYPA, HBGG, D7S8, GC and D1S80. Again, after Bonferroni correction there was no evidence that the population sample deviates from expectations of independence among alleles at the 10 different PCR-based loci. Thus, the allelic frequency data can be used in human identity testing to estimate the frequency of a multiple PCR-based DNA profile in the Swiss population.

We developed the allele-specific TaqMan polymerase chain reaction (AS-TaqMan PCR) and SYBR Green PCR assays for detecting glycophorin A (GYPA), low-density lipoprotein receptor (LDLR), hemoglobin G (HBGG), D7S8 and group-specific component (GC) alleles. We improved the specificity of detecting a nucleotide substitution by introducing tile additional mismatches at position 2 (3 in GYPA). The differences between threshold cycle (0) values of different genotypes on each of the loci were statistically significantly different. All the genotypes agreed with the results using the AmpliType PM+DQAl PCR Amplification and Typing kit. The AS-TaqMan PCR and SYBR Green PCR assays are simple, rapid, and accurate, as well as suitable for high-throughput applications in a forensic investigation. (C) 2003 Elsevier Science B.V. All rights reserved.