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Quantitative Analysis of mRNA Levels in Xenopus Embryos by Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR)

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Steinbach,  OC
Rupp Group, Friedrich Miescher Laboratory, Max Planck Society;

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Rupp,  RAW       
Rupp Group, Friedrich Miescher Laboratory, Max Planck Society;

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Citation

Steinbach, O., & Rupp, R. (1999). Quantitative Analysis of mRNA Levels in Xenopus Embryos by Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR). In M. Guille (Ed.), Molecular Methods in Developmental Biology: Xenopus and Zebrafish (pp. 41-56). Totowa, NJ, USA: Humana Press.


Cite as: https://hdl.handle.net/21.11116/0000-000D-1E39-3
Abstract
Over the last few years, RT-PCR (1,2) has become a widely accepted method for quantitation of steady-state mRNA levels, particularly in Xenopus. Its unmatched sensitivity and swiftness allows for a high sample throughput with minimal amounts of starting material—considerable advantages over the conventional methods of Northern blotting or RNase protection. Initially, the use of RT-PCR for quantitative analysis was viewed skeptically. This was based on the concern that minor differences in the reaction conditions between samples would erratically influence the exponential rate of PCR amplification; therefore, results would be skewed a priori. This theoretical concern has turned out to be irrelevant for most applications.