Quantitative PCR shows weak fluorescence signal across all wells, making accurate quantification difficult. Signal intensity is lower than expected despite adequate template and reagent quality.
Common Causes
1Use of clear-well plates instead of white wells, causing signal refraction through well walls
2Non-optimal seal clarity (cloudy or tinted films/caps) distorting fluorescence signal transmission to detector
Solutions
1Switch to white-well PCR plates that enhance fluorescence reflection to qPCR detector and reduce signal loss
2Ensure sealing films and caps are optically clear or ultraclear specification to minimize signal distortion