Amplification Plots Dip Below Zero Due to Incorrect Baseline Settings (PCR (Sigma Guide))

Question

Accepted Answer

Amplification plots are clearly abnormal with sections dipping below zero dR; data cannot be used as presented; plots appear distorted Common causes: 1) Inappropriate baseline settings applied by instrument software 2) Automatic baseline detection failed 3) Baseline cycles incorrectly defined Solutions: 1) Examine raw data plot to verify actual amplification profile is normal 2) Manually set appropriate baseline cycles based on raw data (typically the initial linear, baseline phase of the reaction, e.g., cycles 6 to 16) 3) Reanalyze data with corrected baseline settings 4) Consult instrument manual for proper baseline determination