PYROSEQUENCING-BASED BARCODES FOR DYE-FREE MULTIPLEX BIOASSAY

摘要：

We proposed a sequence-based barcoding approach to encode a target with a report base and repeats of two stuffer bases. The barcodes were quantitatively decoded by pyrosequencing without pre-separations, and multiple targets were quantified by peaks of the reporter bases in a pyrogram. Using these barcodes, we have developed a barcoded adapter-based method for multiplex gene-expression analysis; as a proof-of concept, the 12-plex gene expression analysis of four different genes among 12 kinds of tissues of a mouse was successfully demonstrated. In comparison to conventional barcodes based on dyes or nanocrystals, our sequence-based barcodes do not need laser, gel, and electrophoresis. Most importantly, this barcode strategy significantly extends the scope of pyrosequencing capability.

关键词： barcodes pyrosequencing BAMGA gene expression analysis

作者: Chen Zhiyao Zhang Xiaodan Liu Xiqun Zou Bingjie Wu Haiping Li Jinheng Tomoharu Kajiyama Hideki Kambara Zhou Guohua

作者单位: Institute of Molecular Medicine, Huaqiao University, Quanzhou 362021, China Molecular Medicine Engin Huadong Research Institute for Medicine and Biotechnics, Nanjing 210002, China Central Research Laboratory, Hitachi, Ltd. , 1-280 Higashi-Koigakubo, Kokubunji-shi, Tokyo 185-8601, Institute of Molecular Medicine, Huaqiao University, Quanzhou 362021, China Molecular Medicine Engin

会议类型: 国际会议

会议名称: The 7th International Forum on Post-Genome Technologies and China-Japan-Korea Summit on Natural Products(第七届国际后基因组技术论坛及中日韩天然产物论坛 IFPT)

会议地点: 重庆

会议语种:英文

页码: 369-373

在线出版日期: 2011-10-28（万方平台首次上网日期，不代表论文的发表时间）