IN-SITU IOCALIZED ROLLING CIRCLE AMPLIFICATION FOR GENOTYPING SINGLE NUCLEOTIDE POLYMORPHISMS

摘要

Single nucleotide polymorphisms are the foundation of powerful complex trait and pharmacogenomic analyses. Herd a novel method based on in-situ isothermal amplification reaction for discriminating multiple loci from multiple human genomes on a microarray is described. Single nucleotide polymorphisms were discriminated from template DNA by specific ligation of a pair of open circle probes in solution and ligation products were then annealed with an acrylamide-modified common primer. Then the mixture included annealed probes and acrylamide monomer was spotted and then immobilized on the slip by polymerization. Gel pads including different annealed probes were in-situ parallelly amplified on the microarray by rolling circle amplification, and amplification products were detected by dual-color fluorescent dyes. The results indicated that in-situ localized rolling circle amplification on microarrays was feasible and allowed efficient discrimination of alleles from different samples. This method may be a promising robustness assay for detecting numerous target sequences.