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.

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

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) ★ 88

Step-by-Step PCR Experiment Protocol Guide

"Step-by-step PCR demonstration showing tube setup, sealing, and thermal cycling where evaporation/sealing failures occur."

Addgene ★ 85

Polymerase Chain Reaction (PCR) Protocol

"PCR protocol walkthrough from Addgene covers reaction setup and cycling, the exact context for plate/film sealing and evaporation issues."

Bilibili (China-Accessible Mirrors) ★ 72

qPCR Principles, Experimental Workflow and Results Analysis

"qPCR workflow video shows plate/film sealing and cycling steps where sample evaporation and loss can occur."