Abstract
High Coverage gene expression profiling (HiCEP) is an AFLP-based method for transcriptome analysis, which doesn't require any sequence information beforehand and is thus applicable in all species. HiCEP observes more than 70% of the entire transcripts and its signals can be basically assigned to certain transcript with one-to-one relationship. Furthermore, it can discriminate 1.5-fold differences in gene expression.
For spreading HiCEP technology through life science field, the analysis should be easy for everyone and high throughput system of it also should be established. For this purpose, we attempted to develop an automatic reaction robot (HiCEPer) which performs the main body of the HiCEP reaction, from 1st strand cDNA synthesis to adapter ligation. HiCEPer basically consists of three modules, for liquid handling, for incubation and for purification of nucleic acid with magnetic beads, and achieves 96reactions simultaneously with 1ng total RNA, which corresponds to 100 mammalian cells. Thus, the development of HiCEPer enabled us to perform massive and sensitive transcriptome analysis in a large number of applications.
