QUANTITATIVE ANALYSIS OF GENE EXPRESSION IN A SINGLE CELL BY QPCR
A method has been developed for analyzing quantitative gene expressions in a single cell. It features a reusable single-cell cDNA library and uses quantitative polymerase chain reaction (qPCR). The library is constructed by converting almost all the mRNA in a single cell into cDNA immobilized on beads. It can be used repeatedly for analyzing multiple gene expressions by qPCR. This method enables to analyze target cDNA of from several to several hundred thousands copies with an experimental error of 15. 9 ％ at most. The gene expression levels for four housekeeping genes (EEF1G, B2M, SDH A, and TBP) in single cells were successfully analyzed. Although the average numbers of cDNA molecules correlated with those for the pooled cell samples, the expression levels fluctuated greatly from cell to cell, which is far bigger than the error levels.
Single-cell cDNA library magnetic beads gene ezpression qPCR
Kiyomi Taniguchi Tomoharu Kajiyama Hideki Kambara
Hitachi, Ltd., Central Research Laboratory 1-280, Higashi-koigakubo Kokubunji-shi, Tokyo 185-8601, Japan
国际会议
The 6th International Forum on Post-genome Technologies(6IFPT)(第六届国际后基因组生命科学技术学术论坛)
北京
英文
413-417
2009-09-17（万方平台首次上网日期，不代表论文的发表时间）