A reagentless biosensor for mRNA: a new tool to study transcription
作者： Alexander Cook , Yukti Hari-Gupta , Christopher P. Toseland
DOI: 10.1101/142794
关键词: Biosensor 、 Biochemistry 、 Messenger RNA 、 Polymerase 、 Gene expression 、 RNA 、 Fluorescence 、 Nucleic acid 、 Transcription (biology) 、 Biology
摘要: Gene expression, catalysed by RNA polymerases, is one of the most fundamental processes in living cells. Yet, means to study their activity are currently limited. The majority methods quantify mRNA based upon initial purification nucleic acid. This leads experimental inaccuracies and loss product. Here we describe use a reagentless fluorescent biosensor single stranded binding (SSB) protein. In this study, SSB showed similar properties mRNA, that its native substrate, ssDNA. Furthermore, MDCC-SSB gave same fluorescence response with both ssDNA ssRNA, concentration dependent manner. When directly compared RT-qPCR, found be more reproducible no product lost through purification. Therefore, can used as comparative measurement concentrations following vitro transcription.
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