Real-Time PCR Troubleshooting Tool

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Real-Time PCR Troubleshooting Tool

Having problems with your gene expression or SNP genotyping experiments? Do your amplification curves look sigmoidal, or do you have no curves at all? Do your allelic discrimination plots have diffuse or trailing clusters? Our interactive troubleshooting tool will guide you step by step to a solution.

Gene Expression/Quantitation Experiments

I am having problems with...
	Abnormal Amplification Your curves are sigmoidal or amplification occurs later than you expected.
	NTC Positive Amplification You have an amplification product in your no template control (NTC).
	No Amplification You have no detectable PCR product.
	Poor PCR Efficiency If the slope of your standard curve is below -3.6, you have poor PCR efficiency.
Example of a normal amplification curve
	Normal Amplification Curve Your amplification curve should look something like this.

SNP Genotyping Experiments

I am having problems with...
	SNP Not Amplifying You cannot amplify a SNP.
	Trailing Clusters Not using ROX or not designating it as a passive reference dye may cause trailing clusters.
	Diffuse Clusters You have diffuse clustering in your SNP analysis.
	No Autocalling Failure of autocalling of SNP results is usually due to outlier wells or software problems.
Example of a normal allelic discrimination plot
	Normal Allelic Discrimination Plot Your cluster plot should look something like this.