Using the Access RT-PCR System: Reaction Parameters that Affect Efficient Amplification
RT-PCR should be performed in the absence as well as presence of the reverse transcriptase, to assess DNA contamination in the template RNA. In addition, "no-template" negative control reactions should always be performed.
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The analysis of RNA using amplification-based methodologies is a fundamental technique used in many research and diagnostic applications. While the isolation of high-quality RNA is fundamental to the success of RT-PCR, a variety of reaction parameters can profoundly affect efficient amplification of target sequences. This article demonstrates how titration of several amplification reaction components can affect amplification of endogenous RNA targets using the Access RT-PCR System.
Promega Notes 73, 14.
Publication Date: 1999