AccuCleave Agarose

Developed for low-throughput, agarose-based laboratories, AccuCleave – Agarose provides uncompromising detection of CRISPR/Cas9 gene editing events.

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DNA-100-0100

DNA-100-0100 AccuCleave™ T7

AccuCleave Agarose Kits
100 Sample AccuCleave T7 Kit

Streamline your CRISPR workflow with the AccuCleave T7 Kit. A meticulously calibrated and rigorously tested heteroduplex cleavage assay, that enables the quick and accurate identification of gene editing events. Optimized for agarose electrophoresis, the AccuCleave T7 Kit works with any thermocycler and agarose electrophoresis set up.

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Components

Components 100 Sample Kit
T7 Endonuclease 8 µL (CP-END)
10X Cleavage Buffer 1.1 mL (CP-CVB)
WT Control 20 µL (CP-PC1-U020)
-2 bp Control 20 µL (CP-PC5-U020)
Primer Set 1 20 µL (CP-PRM1-U020)

Examples

Efficient CRISPR gene editing workflows necessitate the use of an accurate screening protocol in order to minimize the cost of downstream characterization procedures. Researchers are able to easily visualize results using agarose gel electrophoresis, shown in Figure 1.

Figure 1
Figure 1. CRISPR event detection example. Control Fragments were amplified using the following ratios and analyzed with or without digestion by T7 Endonuclease. Lanes 2 and 3 WT/-2 bp (9.5/0.5 m/m), Lanes 4 and 5 WT/ -2 bp/-2 bp (50/50 m/m), Lanes 6 and 7 WT/-2 bp/+2 bp (50/25/25 m/m/m).

Percent cleaved can be estimated using image processing software such as ImageJ to obtain relative quantification of amplicons and cleavage products. Once relative quantification values are obtained, percent cleaved can be estimated using the following equation:

Figure 2

AATI has simplified determination of percent cleaved with a formula that uses values directly from ImageJ.

Further information about a positive clone, including the zygosity and the exact nature of the editing event, must be determined by NGS or Sanger Sequencing.

Additional Resources

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