T7E1说明书(NEB).pdfVIP

T7E1 protocol 1. CRISPR Transfection. 48~72h 2. Genomic DNA prep. 3. Target site PCR. 4. Denature/Reannealing 5. T7 endonuclease I reaction. 6. Agarose gel eletrophoresis T7E1 protocol 1. CRISPR Transfection.  Methods and conditions for transfection varies for each 48~72h cell lines. Use the optimized transfection protocol (reagents, DNA amount, Etc.) for your cell line !! Caution Optimal transfection efficiency (at least 50%) is critical. - Nucleofection seems to be most broadly applicable. T7E1 protocol 48~72h 2. Genomic DNA prep.  Genomic DNA prep at 48-72 hours post-transfection using any commercial or in-house genomic DNA prep protocol/kit T7E1 protocol 48~72h Most important step!  Robust and clean PCR amplification is critical. Thus,