CRISPR-mediated Editing Detection

CRISPR editing detection and quantification by T7 assay, sequencing, and ddPCR

After CRISPR editing, you need to confirm and quantify what happened. This section covers T7 endonuclease (Surveyor or T7E1) digestion, Sanger sequencing with TIDE or ICE analysis, deep amplicon sequencing, and ddPCR-based quantification.

Pick by quantitative resolution and cost.

1. Fast semi-quantitative. Choose T7E1 digestion. Fastest and cheapest. Resolves indels but does not quantify clonal allele frequencies.
2. Quantitative at moderate cost. Choose Sanger sequencing plus TIDE or ICE analysis. The cost-quality sweet spot for routine work.
3. Highest precision. Choose deep amplicon sequencing. Most quantitative but most expensive. Useful for clonal-level work.
4. Edit type. Small indels versus large deletions versus precise knock-ins each have suitable methods.
5. Throughput. T7E1 single-by-single. TIDE in modest batches. Amplicon-seq in pooled batches.

ABM supplies editing-detection kits in this inventory.