National Taiwan Ocean University Institutional Repository:Item 987654321/41336
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题名: Verifying expressed transcript variants by detecting and assembling stretches of consecutive exons
作者: Tzu-Hung Hsiao;Chien-Hong Lin;Te-Tsui Lee;Ji-Yen Cheng;Pei-Kuen Wei;Eric Y. Chuang;Konan Peck
贡献者: 國立臺灣海洋大學:機械與機電工學系
关键词: polymerase chain reaction;electrophoresis;capillary;exons;genes;rna;splicing;software;blood capillaries;amplification
日期: 2010-08
上传时间: 2017-02-14T07:03:45Z
摘要: Abstract:We herein describe an integrated system for the high-throughput analysis of splicing events and the identification of transcript variants. The system resolves individual splicing events and elucidates transcript variants via a pipeline that combines aspects such as bioinformatic analysis, high-throughput transcript variant amplification, and high-resolution capillary electrophoresis. For the 14 369 human genes known to have transcript variants, minimal primer sets were designed to amplify all transcript variants and examine all splicing events; these have been archived in the ASprimerDB database, which is newly described herein. A high-throughput thermocycler, dubbed GenTank, was developed to simultaneously perform thousands of PCR amplifications. Following the resolution of the various amplicons by capillary gel electrophoresis, two new computer programs, AmpliconViewer and VariantAssembler, may be used to analyze the splicing events, assemble the consecutive exons embodied by the PCR amplicons, and distinguish expressed versus putative transcript variants. This novel system not only facilitates the validation of putative transcript variants and the detection of novel transcript variants, it also semi-quantitatively measures the transcript variant expression levels of each gene. To demonstrate the system’s capability, we used it to resolve transcript variants yielded by single and multiple splicing events, and to decipher the exon connectivity of long transcripts.
URI: http://ntour.ntou.edu.tw:8080/ir/handle/987654321/41336
显示于类别:[機械與機電工程學系] 期刊論文

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