Part:BBa_K1473008:Experience

We put a sacI restriction cutting site downstream to gRNA (not shown in sequence information) and ligated it back to pSB1C3 with correct prefix and surffix. Then we cut this recombinant plasmid with sacI and run gel electrophoresis for testing. As we can see in this image, because pSB1C3 only contains 1 sacI cutting site itself, so it would not be cut into 2 parts when no gRNA is ligated inside. But if there is, then sacI will cut this recombinant plasmid into 2 parts, whose lengths are 939 bp and 1159 bp. Obviously, we have successfully synthesized this gRNA.