In CRISPR pooled screening, the quality of your starting sgRNA library directly determines whether your hit list reflects true biology or technical bias. A library with poor coverage or uneven representation can silently drop essential targets before the screen even begins.
 
We recently completed a genome-wide sgRNA library project for a leading cancer research team studying drug resistance. Their requirements were stringent:
  • 100× clone coverage
  • 70% correct reads (on‑target sequences)
  • 99% NGS‑verified library coverage
  • Skew ratio <5
Our final delivery exceeded every metric:
  • Clone coverage met the >100× threshold.
  • Correctness rate was well above 70%.
  • NGS coverage reached 99.92% - meaning nearly every sgRNA was detected, minimising the risk of missing critical targets.
  • Skew ratio was 3.2 -far below 5, indicating exceptionally even abundance across all sgRNAs, which greatly reduces false negatives during viral transduction and cell passaging.
          
All plasmids were prepared with endotoxin-free maxiprep and validated by deep NGS at multiple stages for sequence accuracy and uniform representation.
Questions on sgRNA libraries or quality?


 
 
 
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