Home › Failure Case Library › PCR Sample Evaporation and Loss During Cycling
PCR / qPCR Plastics severe

PCR Sample Evaporation and Loss During Cycling

Symptom
Visible reduction in reaction volume after thermal cycling, with condensation on tube caps or film. May result in concentrated reagents, failed reactions, or inability to recover product.
Common Causes
  1. 1 PCR film not fully sealing wells, particularly along outer edges and between wells
  2. 2 Raised deck plates or raised lettering preventing complete film contact during automatic sealing
  3. 3 Cap strips not firmly seated or incompatible with tubes/plates being used
Solutions
  1. 1 Press film firmly along every edge (including outer edges) and between/around wells; use sealing applicator tool for uniform pressure
  2. 2 Choose plates with flat deck (not raised) and engraved lettering (not raised) to facilitate complete sealing, especially for automation
  3. 3 Verify cap strip compatibility and ensure tightly in place using cap installing tool
Related Video (3)
Bilibili (China-Accessible Mirrors) ★ 85
Step-by-Step PCR Experiment Protocol Guide
"Comprehensive hands-on PCR protocol demonstration covering complete workflow including proper setup and handling techniques that directly address sealing and sample retention issues."
Addgene ★ 80
Polymerase Chain Reaction (PCR) Protocol
"PCR protocol walkthrough by Addgene showing proper technique and reagent handling, providing foundational context for understanding where evaporation failures occur during thermal cycling."
Bilibili (China-Accessible Mirrors) ★ 72
qPCR Principles, Experimental Workflow and Results Analysis
"qPCR experimental workflow demonstration with detailed protocol steps that includes equipment handling and setup procedures relevant to preventing condensation and volume loss."
Source: thermofisher.com ↗
← Back to all cases