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) ★ 78

Step-by-Step PCR Experiment Protocol Guide

"Hands-on PCR workflow demonstration covers tube/film setup and thermal cycling where sealing failures and evaporation occur."

Addgene ★ 75

Polymerase Chain Reaction (PCR) Protocol

"Detailed PCR protocol walkthrough shows correct reaction setup and sealing steps a researcher can follow to avoid evaporation."

Bilibili (China-Accessible Mirrors) ★ 70

qPCR Principles, Experimental Workflow and Results Analysis

"qPCR protocol demonstration covers plate/film sealing and cycling context relevant to diagnosing volume loss."