TITLE

Comparison of PrASE and Pyrosequencing for SNP Genotyping

AUTHOR(S)
Kaller, Max; Hultin, Emilie; Holmberg, Kristina; Persson, Marie-Louise; Odeberg, Jacob; Lundeberg, Joakim; Ahmadian, Afshin
PUB. DATE
January 2006
SOURCE
BMC Genomics;2006, Vol. 7, p291
SOURCE TYPE
Academic Journal
DOC. TYPE
Article
ABSTRACT
Background: There is an imperative need for SNP genotyping technologies that are cost-effective per sample with retained high accuracy, throughput and flexibility. We have developed a microarray-based technique and compared it to Pyrosequencing. In the protease-mediated allelespecific extension (PrASE), the protease constrains the elongation reaction and thus prevents incorrect nucleotide incorporation to mismatched 3'-termini primers. Results: The assay is automated for 48 genotyping reactions in parallel followed by a tagmicroarray detection system. A script automatically visualizes the results in cluster diagrams and assigns the genotypes. Ten polymorphic positions suggested as prothrombotic genetic variations were analyzed with Pyrosequencing and PrASE technologies in 442 samples and 99.8 % concordance was achieved. In addition to accuracy, the robustness and reproducibility of the technique has been investigated. Conclusion: The results of this study strongly indicate that the PrASE technology can offer significant improvements in terms of accuracy and robustness and thereof increased number of typeable SNPs.
ACCESSION #
28858730

 

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