Real-Time PCR Troubleshooting

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Overview

Are you having trouble with your real-time PCR experiment? Do you need help diagnosing your experiment? The real-time PCR doctor is here to help. Unexpected fluorescence data are symptomatic of problems with your real-time PCR reaction components or amplification protocol. Click one of the symptoms below to learn about possible causes and treatments.

Related Topics: What Is Real-Time PCR?, How Real-Time PCR Works, and qPCR Assay Design and Optimization.

 

 

 



Allelic Discrimination Experiments

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High Resolution Melt (HRM) Experiments:

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If you don't see your symptom listed above, please contact technical support at 1-800-424-6723, option 2.


Further Reading

Hu CJ et al. (2006). Differential regulation of the transcriptional activities of hypoxia-inducible factor 1 alpha (HIF-1alpha) and HIF-2alpha in stem cells. Mol Cell Biol 26, 3514–3526. PMID: 16611993

Ivanova N et al. (2006). Dissecting self-renewal in stem cells with RNA interference. Nature 442, 533–538. PMID: 16767105

Li Y et al. (2006). DAPK1 variants are associated with Alzheimer's disease and allele-specific expression. Hum Mol Genet 15, 2560–2568. PMID: 16847012

Morozumi M et al. (2006). Simultaneous detection of pathogens in clinical samples from patients with community-acquired pneumonia by real-time PCR with pathogen-specific molecular beacon probes. J Clin Microbiol. 44, 1440–1446. PMID: 16597874

Ridd K et al. (2006). Association of gene expression with sequential proliferation, differentiation, and tumor formation in murine skin. Carcinogenesis 27, 1556–1566. PMID: 16537558

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Literature
Number Description Download
5692 Real-Time qPCR as a Tool for Evaluating RNAi-Mediated Gene Silencing, Rev A (tech note) Click to download
5962 Fast Optimization of a Multiplex Influenza Identification Panel Using a Thermal Gradient, Rev A Click to download
6004 A Practical Guide to High Resolution Melt Analysis Genotyping, Rev A Click to download
6009 High Resolution Melt Parameter Considerations for Optimal Data Resolution, Rev A Click to download
6047 Optical Design of CFX96 Real-Time PCR Detection System Eliminates the Requirement of a Passive Reference Dye, Rev A Click to download
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