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Experiments & Protocols

Experiments

Protocols

BsaI is a restriction enzyme with unique type 2 properties. What this means is that a sequence of DNA can be designed to have a desired overhang on each side. The BsaI will cut itself out of the sequence, as shown in the picture below:

After cutting its recognition sequence out and leaving the desired overhang, a gRNA with matching sticky ends can be inserted into the space. In other words, the gRNA will completely take the place of the BsaI restriction site.