![]() It is also demonstrated here that high genotyping throughput can be achieved by combining this method with the microplate array diagonal gel electrophoresis (MADGE) technique ( 12). Rules for selecting a nucleotide for the additional mismatch in classical ARMS PCR have been described previously ( 8): a ‘strong’ mismatch (G/A or C/T mismatches) at the 3′-terminus of an allele-specific primer will likely require a ‘weak’ second mismatch (C/A or G/T) and vice versa, whereas a ‘medium’ mismatch (A/A, C/C, G/G or T/T) at the 3′-terminus will likely require a ‘medium’ second mismatch. An extra destabilizing mismatch has been found to increase the specificity of classical ARMS-PCR ( 6, 8– 11). In contrast to Bi-PASA, both inner primers of the tetra-primer ARMS-PCR method encompass a deliberate mismatch at position –2 from the 3′-terminus. Differences between the tetra-primer ARMS-PCR method, the original tetra-primer PCR method and the Bi-PASA (bidirectional PCR amplification of specific alleles) method reported by Liu et al. The technique, named tetra-primer ARMS-PCR, adopts certain principles of the tetra-primer PCR method ( 5) and the amplification refractory mutation system (ARMS) ( 6 Fig. Developing such techniques has thus become one of the new goals of the Human Genome Project ( 4).Ī simple and economical SNP genotyping method involving a single PCR reaction followed by gel electrophoresis is reported here. Because such studies will involve analysis of large marker sets in large numbers of subjects, they will benefit from rapid, simple, low cost and high throughput methods for SNP genotyping. The high density and mutational stability of SNPs make them particularly useful DNA markers for population genetics and for mapping susceptibility genes for complex diseases ( 3). ![]() ![]() It is estimated that single nucleotide polymorphisms (SNPs) occur on average once per 250–1000 bp and account for ∼90% of DNA sequence variants in the human genome ( 1, 2). Received FebruRevised Accepted July 12, 2001. ![]()
0 Comments
Leave a Reply. |
AuthorWrite something about yourself. No need to be fancy, just an overview. ArchivesCategories |