IN-SITU IOCALIZED ROLLING CIRCLE AMPLIFICATION FOR GENOTYPING SINGLE NUCLEOTIDE POLYMORPHISMS
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.
DNA microarray in-situ RCA ligation polyacrylanude gel single nucleotide polymorphisms
Xiujie Li Pengfeng Xiao Zuhong Lu
State Key Laboratory of Bioelectronics, Southeast University,Nanjing 210096, China
国际会议
The 4th International Forum on Post-genome Technologies(4IFPT)(第四届国际后基因组生命科学技术学术论坛)
杭州
英文
130-133
2006-09-25（万方平台首次上网日期，不代表论文的发表时间）