2002
DOI: 10.1093/nar/gnf074
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SNP typing by apyrase-mediated allele-specific primer extension on DNA microarrays

Abstract: This study reports the development of a microarray-based allele-specific extension method for typing of single nucleotide polymorphisms (SNPs). The use of allele-specific primers has been employed previously to identify single base variations but it is acknowledged that certain mismatches are not refractory to extension. Here we have overcome this limitation by introducing apyrase, a nucleotide-degrading enzyme, to the extension reaction. We have shown previously that DNA polymerases exhibit slower reaction ki… Show more

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Cited by 50 publications
(21 citation statements)
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“…Cluster or scatter plot analysis has become the method of choice for genotype assignment and determination of cutoff values for the three allelic genotypes (13,27,33,34). Tight clusters indicate good genotype discrimination, and the larger the distance between the clusters the better is the discrimination between homozygous and heterozygous genotypes (27).…”
Section: Resultsmentioning
confidence: 99%
“…Cluster or scatter plot analysis has become the method of choice for genotype assignment and determination of cutoff values for the three allelic genotypes (13,27,33,34). Tight clusters indicate good genotype discrimination, and the larger the distance between the clusters the better is the discrimination between homozygous and heterozygous genotypes (27).…”
Section: Resultsmentioning
confidence: 99%
“…9). The designed microfluidic confined bead array was successfully demonstrated for apyrase assisted allele-specific primer extension [20,82]. This approach showed reliable discrimination of synthetic target DNA of 0.5 pM concentration that corresponded to an absolute detection sensitivity of approximately one attomole of target molecules in a 15 µL sample.…”
Section: Microfluidic Arraysmentioning
confidence: 99%
“…Over the past twenty years, many different methods have been developed for SNP genotyping by PCR, including hybridization 16 , allele-specific PCR 17 , primer extension 18 , oligonucleotide ligation 19 , direct DNA sequencing 20 and endonuclease cleavage after amplification of the subjected genomic region by PCR [21][22] . Recent technologies for SNP genotyping (Taq Man method [23][24] , Invader method [25][26] , MALDI-TOF method 27 , GeneChips 28 ) have the potential for high-throughput, but they require the purchase of expensive equipment.…”
Section: Introductionmentioning
confidence: 99%