qPCR Primer Design: NCBI Screencast Tutorial

severe

No Band Due to Primer Design or Quality Issues

No PCR product is obtained despite correct thermal cycling and component concentrations. Investigation reveals potential primer-related issues including design flaws or contamination.

💡 5 · ✓ 5

severe

Insensitive Assay with Abnormal Amplification Due to Probe Secondary Structure

Assay is insensitive and amplification plots look abnormal with pronounced drift of the baseline; fluorescence signals are weak or irregular

💡 3 · ✓ 4

severe

No DNA Amplification in ChIP Samples

No PCR product is detected in ChIP samples by qPCR or gel electrophoresis. Ct values fail to appear or remain above threshold, and no visible bands are observed on gels.

💡 5 · ✓ 6