Home Resources Troubleshooting Guides Troubleshooting Guide for use with SARS-CoV-2 Rapid Colorimetric LAMP Assay Kit (NEB #E2019)

Troubleshooting Guide for use with SARS-CoV-2 Rapid Colorimetric LAMP Assay Kit (NEB #E2019)

Note: For additional assistance please refer to the product FAQ

PROBLEM POSSIBLE CAUSE(S) SOLUTION(S)
Colorimetric LAMP reaction is orange prior to amplification Nucleic acid sample input is incompatible
  • Dilute sample in nuclease- free water or adjust the pH to ~8.0 prior to addition to the reaction
Repeated exposure of the colorimetric LAMP master mix to atmosphere causes a slight pH drop
  • Avoid extended air exposure by closing the cap and storing the master mix in the vial provided at -20°C
Positive Control reaction fails to turn yellow following incubation at 65°C Improper pipetting during colorimetric LAMP set-up
  • Ensure proper pipetting techniques are used.
Poor mixing of reagents during set-up
  • Make sure all reagents are properly mixed after thawing
  • Ensure reactions are properly mixed prior to incubation
NTC reaction turns yellow following incubation at 65°C   Reagents are contaminated with the positive control
  • Replace all stocks and reagents
  • Clean equipment and setup area with 10% chlorine bleach
Improper pipetting during colorimetric LAMP set-up
  • Ensure proper pipetting techniques are used
Internal Control reaction fails to turn yellow following incubation at 65°C Improper pipetting during colorimetric LAMP set-up
  • Ensure proper pipetting techniques are used
Sample nucleic acid is incompatible with colorimetric LAMP
  • Dilute sample in nuclease-free water or adjust the pH to ~8.0 prior to addition to the reaction
Sample nucleic acid is isolated from a non-human source
  • Use human nucleic acid input
  • Design LAMP primers for your specific sample source
Poor mixing of reagents during set-up
  • Make sure all reagents are properly mixed after thawing
  • Ensure reactions are properly mixed prior to incubation
RNA template is contaminated or degraded
  • Prepare high quality RNA