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- W1964426538 abstract "A novel multiplexing method, which relies on universal amplification of separated ligation-dependent probes (ASLP), has been developed to genotype single-nucleotide polymorphisms (SNPs). The ASLP technique employs two allele-specific oligonucleotides (ASO), modified with universal forward primer sequences at the 5′-end and a common locus-specific oligonucleotide (LSO) extended with a universal separation (US) sequence at the 3′-end. In the process, allele-specific ligation first takes place when target genomic DNA is hybridized by perfectly matching the ASO together with the LSO. A separation probe, which consists of a universal reverse primer sequence labeled with biotin at the 5′-end and complementary sequence of US at the 3′-end, is then applied to the resulting ligation product. During the extension reaction of the separation probe, the ligated probes dissociate from target genomic DNA in the form of a double-stranded DNA and are separated from the reaction mixture, which includes genomic DNA and unligated probes, by simply using streptavidin-coated magnetic beads. PCR amplification of the separated ligation products is then carried out by using universal primers and the PCR products are hybridized on a DNA microarray using the RecA protein. The advantageous features of the new method were demonstrated by using it to genotype 15 SNP markers for cultivar identification of pepper in a convenient and correct manner." @default.
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- W1964426538 date "2014-04-01" @default.
- W1964426538 modified "2023-09-26" @default.
- W1964426538 title "Application of the ASLP technology to a novel platform for rapid and noise-free multiplexed SNP genotyping" @default.
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- W1964426538 doi "https://doi.org/10.1016/j.bios.2013.10.071" @default.
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