Real-Time PCR Troubleshooting

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

Back to Top
 

High Resolution Melt (HRM) Experiments:

Back to Top
 

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

Page Contents

Videos

Quantitative PCR (qPCR): Validating a SYBR® Green I Assay
This tutorial describes the characteristics of an optimized SYBR Green I quantitative PCR (qPCR) assay. qPCR assays must be optimized to ensure results that are biologically and statistically significant. Topics include a brief review of qPCR chemistry, with an emphasis on SYBR Green I reactions, and definitions of the four main characteristics, or hallmarks, of an optimized qPCR assay.
 
Quantitative PCR (qPCR): Hallmarks of an Optimized SYBR Green I Assay
This tutorial describes the characteristics of an optimized SYBR® Green I quantitative PCR (qPCR) assay. qPCR assays must be optimized to ensure results that are biologically and statistically significant. Topics include a brief review of qPCR chemistry, with an emphasis on SYBR Green I reactions, and definitions of the four main characteristics, or hallmarks, of an optimized qPCR assay.
 
Data Analysis: Real-Time Quantitative PCR
This tutorial pertains to the analysis of real-time quantitative PCR (qPCR) data.
 

Documents MADHA

Number Description Options
7216
End the Cycle of Repeated, Failed Experiments with a Robust Field Guide to qPCR
6894
Tips, Tricks & Best Practices: The Ultimate qPCR Assay Design Guide
[ Add to Cart (Free) ]
5692
Real-Time qPCR as a Tool for Evaluating RNAi-Mediated Gene Silencing, Rev A
6047
Optical Design of CFX96™ Real-Time PCR Detection System Eliminates the Requirement of a Passive Reference Dye, Rev A
5962
Fast Optimization of a Multiplex Influenza Identification Panel Using a Thermal Gradient, Rev A