Multiplexed genotyping of single nucleotide polymorphisms using microarray technology

This thesis was composed of four separate studies. The first study was initiated to adapt minisequencing to solid support to interrogate multiple SNPs which had been reported to be associated with coronary artery disease. This microarray-based platform has demonstrated a great potential for highly parallel genotyping. Secondly, an innovative two-color platform (TATLIPEXA) was successfully developed which could accurately interrogate all kinds of SNPs and some short insertions/deletions simultaneously with the use of only two fluorescently labelled ddNTPs. Thirdly, by exploiting the unique feature of TdT to polymerize oligonucleotides in a template-independent manner, another novel and versatile tool (TAPE) was established and it demonstrated great potential in genomic analyses, mainly genotyping and gene expression. Finally, an unusual property of TdT was discovered during the effort to develop TAPE, which directly challenged a common belief that 3' chemically modified oligonucleotides could not be polymerized because the 3'-OH was occupied by appropriate modifier.

Author dc:creator

• LI JINGGUANG

• single nucleotide polymorphisms, minisequencing, tag-anti-tag liquid-phase primer extension array, TdT-assisted probe elongation