detection of genetic variations in coagulopathy-related genes using ramified rolling circle amplification
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2014
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Abstract
We evaluated single nucleotide polymorphism (SNP) detection via a target-capture, C-probe ligation, and RAM assay in a single-blind comparison to clinical samples that had been tested with FDA-cleared tests for up to 4 different vascular disease-related SNPs. In the RAM assay circulizable linear probes (C- or padlock probes) were annealed directly to genomic DNA, processed on a largely automated platform, and ligated C-probes were amplified by real-time RAM. After allele determinations were made with the experimental system, the sample genotypes were unblinded and the experimentally determined genotypes were found to be completely consistent with the FDA-cleared test results. The methods and results presented here show that a combination of C-probes, automated sample processing, and isothermal RAM provides a robust, and specific, nucleic acid detection platform that is compatible with automated DNA sample preparation and the throughput requirements of the clinical laboratory.
| Reference Key |
smith2014biomeddetection
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| Authors | ;James H. Smith;Miao Cui;David Y. Zhang;Thomas P. Beals;Fei Ye |
| Journal | spectrochimica acta - part a: molecular and biomolecular spectroscopy |
| Year | 2014 |
| DOI |
10.1155/2014/641090
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