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T7E1 Assay Protocol

t7e1 assay protocol 01Step1:CRISPR Delivery

Delivery methods:

Lentivirus Transduction

Plasmid transfection

Ribonucleoprotein delivery

t7e1 assay protocol 02Step2:Genome Extraction

Please following the procedure in Genomic Extraction Kit

Or using the Direct PCR Kit to treat cell samples

t7e1 assay protocol 03 Step3:Nest PCR for Target site

Nest PCR

First round PCR: 20 cycles

800-1000bp length

Second round PCR: 30-40cycles

Around 500bp length

t7e1 assay protocol 04 Step4:PCR product Annealing

Annealing procedure

t7e1 assay protocol 07

t7e1 assay protocol 05 Step5:T7E1 treatment

Please following the procedure in Knockout Detection Kit

Or using T7E1 enzyme for incubation in 37℃ for 30min

t7e1 assay protocol 06 Step6:Gel electrophoresis

Data shall be visualized by 1.2%-1.5% agarose gel

Note: direct PCR product could be used for T7E1 detection.

After treated with T7E1, reactions should be electrophoresed by high concentration agarose gels.

For research use only. Not intended for any clinical use.

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