Internal Control Fails Due to RNA Template Degradation

Symptom

The internal control reaction fails to amplify when using RNA samples, while DNA-based controls work normally, indicating RNA-specific quality issues.

1. 1 RNA template degraded by endogenous or environmental RNases during sample collection or storage
2. 2 Improper RNA storage conditions (e.g., storage at -20°C instead of -80°C, multiple freeze-thaw cycles)
3. 3 RNase contamination from skin, dust, or laboratory equipment during sample handling
4. 4 RNA extraction performed without adequate RNase inhibition or inadequate washing steps
5. 5 Low RNA integrity number (RIN <7.0) due to tissue autolysis or delayed processing

1. 1 Prepare high-quality RNA using RNase-free technique: wear gloves, use dedicated RNase-free pipettes and tips
2. 2 Store RNA samples at -80°C in single-use aliquots to avoid freeze-thaw degradation
3. 3 Use RNA extraction kits with integrated RNase inhibitors and follow manufacturer protocols precisely
4. 4 Treat all plasticware and surfaces with RNase decontamination solution (e.g., RNaseZap) before RNA work
5. 5 Process samples immediately after collection or stabilize in RNA preservation reagent (e.g., RNAlater)
6. 6 Assess RNA quality by gel electrophoresis or Bioanalyzer before LAMP; use only samples with intact 18S/28S rRNA bands