ddPCR Genome Edit Detection Assays

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Overview

ddPCR Genome Edit Detection Assays offer a fast, precise, simple, and cost-effective method for detection of genome editing events created using nucleases that cause double-stranded breaks in the DNA such as CRISPR-Cas9. Assays to detect HDR (Homology Directed Repair) and NHEJ events (Non-homologous end joining events) can be obtained for any target using Bio-Rad’s easy-to-use, intuitive Digital Assay site.

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ddPCR Genome Edit Detection Assays offer a fast, precise, simple, and cost-effective method for detection of genome editing events created using nucleases that cause double-stranded breaks in the DNA such as CRISPR-Cas9. Assays to detect HDR (Homology Directed Repair) and NHEJ events (Non-homologous end joining events) can be obtained for any target using Bio-Rad’s easy-to-use, intuitive Digital Assay site.

Droplet Digital PCR offers a superior ultrasensitive, rapid, and low-cost method for absolute quantification of genome editing events, from very small amounts of starting material.

Key Benefits

• Enable rapid assessment of HDR and NHEJ edits generated by CRISPR-Cas9 or other genome editing tools
• Detect editing events present at frequencies ≤ 0.5%
• Provide absolute quantification of genome editing events from as little as 5ng of total gDNA (cells from a single well of a 96 well plate)
• Distinguish between homozygous and heterozygous edits in clonal populations
• Results are easily analyzed using QX Manager Software

Sample Data

 

Fig. 1. HDR assay readout.HDR mutation positive control (WT + 20% gblocks gene fragment containing a single base pair substitution) with HDR GED + HDR REF Assay. The HDR edits are FAM+HEX-, while the HEX+ and FAM+HEX+ positive clusters represent the total number of copies, including WT, NHEJ, and HDR edits.

 

 

Fig. 2. NHEJ assay readout.NHEJ mutation-positive control (WT + 1% gblocks gene fragment containing a one base pair deletion at the predicted cut site). The WT cluster is positive for both FAM and HEX, while the NHEJ mutant single-positive droplets are positive for FAM only.

 

Packaging Options

Genome edit detection assays, optimized specifically for ddPCR, can be ordered instantly for any target of interest on Bio-Rad’s easy to use Digital Assay Site.

Catalog# Product Name Reaction Size
12002314 ddPCR NHEJ Gene Edit Assay, 100 rxns 100
12002315 ddPCR NHEJ Gene Edit Assay, 500 rxns 500
12003794 ddPCR NHEJ Gene Edit Pkg, 1000 rxns 1000*

*Sequences of primers and probes provided

Catalog# Product Name Reaction Size
12002312 ddPCR HDR Gene Edit Assay, 100 rxns 100
12002313 ddPCR HDR Gene Edit Assay, 500 rxns 500
12003796 ddPCR HDR Gene Edit Pkg, 1000 rxns 1000*

*Sequences of primers and probes provided

Catalog# Product Name Reaction Size
12003805 ddPCR HDR Ref Assay, Predesigned, 100rxns 100
12003806 ddPCR HDR Ref Assay, Predesigned, 500rxns 500
12003793 ddPCR HDR Ref Pkg, Predesigned, 1000rxns 1000*

*Sequences of primers and probes provided

Related Products

Resources

Documents

Documents

Number Description Options
7155
Direct Quantification of Genome Editing Efficiency from Whole Cells Using SingleShot Cell Lysis Buffer and ddPCR Genome Edit Detection Assays
10000065278
ddPCR™ NHEJ Genome Edit Detection Assays, Ver A
10000065281
ddPCR™ HDR Genome Edit Detection Assays, Ver A
10000065279
ddPCR™ HDR Genome Edit Detection Assays, Predesigned, Ver A