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生化實驗英翻中
2008/05/16 07:29
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生化實驗英翻中

原文:

We have demonstrated on-chip RNA isolation in picoliter droplets, reverse transcription within the individual droplets, and subsequent real-time PCR with fluorescence detection of amplification in each interrogated droplet. The system employed a method of sample partitioning into monodisperse picoliter droplets emulsified in oil to generate isolated chemical reactors for highsensitivity nucleic acid detection. The method required 23 cycles for single-copy real-time reverse transcription from RNA, amplification, and detection on-chip using Taqman-based probes. The results show the method is well-suited to quantitative PCR applications, given the observability of Poisson statistics in the picodroplet-discretized sample. Applying digital microfluidics to real-time RT-PCR combines the advantages of on-chip processing of picoliter reactors with single-copy detection.

 

譯文:

我們已說明在picoliter(10-12)的滴液RNA切片的隔離,在各別的滴液中反轉錄,然後在各測試滴液中,使用螢光放大測定進行即時PCR處理。該系統使用將樣本分離單分散性彌散體的picoliter大小的滴液,在油液中乳化的方法,為高敏感度的核酸偵測,建立隔離的化學性反應子。本方法需使用Taqman探針,進行23次的切片的RNA放大偵測的單複製即時反轉錄處理。結果顯示本方法適用於PCR的量化測定,可用於Pico滴液辨識樣本,在Poisson統計測定的可觀察性。使用數位微液的即時RT-PCR處理,可整合使用單複製偵測Pico尺寸化學反應子切片處理的優點。
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