Gene expression analysis

pcr

RT-qPCR (Reverse transcription quantitative polymerase chain reaction) is the most common approach for quantitation of RNA levels, and this method is widely used in our laboratory.

The method is based on a combination of three steps: 1) The reverse transcription of RNA into complemantary DNA (cDNA), 2) amplification of cDNA of interest using PCR and 3) detection and quantitation of amplified PCR product at each PCR cycle using fluorescent reporter molecules.

We have used RT-qPCR to examine the expression pattern of selected genes in the Serca2 knockout (KO) mice. The figure shows an amplification plot for Serca2 of samples from the control group and the KO group 6 days after disruption of the serca2 gene. The control group, with relative higher expression of the Serca2 gene, creates a detectable PCR-product at an earlier time point (lower cycle number) than the KO group, confirming the downregulation of serca2 in the KO group.

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