SnapGene shows by default only cutters that cut only once in the sequence and that have a restriction site of at least 6nt (red). Go to the BamHI page: there you'll see a lot of useful info on the use of the enzyme together with a list of enzymes with the same restriction site (red) if there are any and a list of enzymes generating compatible sticky ends (green).Ĭheck which enzymes cut only once in the Rab5 PCR product This opens the Restriction Enzymes window. finding compatible enzymesĬheck which enzymes generate sticky ends that are compatible with BamHIĮxpand Enzymes in the top menu and select Restriction Enzymes (red): It has useful features some of them are not avaliable in CLC e.g. SnapGene can help you choosing the appropriate enzymes. You can see the ends that are created by these enzymes by hovering your mouse over their name: Which overhangs are created by these enzymes ? To this end we want to use restriction enzymes from the multiple cloning site in the vector that cut in the primers but not in the gene of interest.įind two enzymes from the multiple cloning site in the vector that cut in the primers but not in the Rab5 gene.īy looking at the enzymes we can see that BspE1 and BamHI cut in the multi-cloning site of the vector and in the primers but not in the Rab5 gene.
We want to clone the amplified Rab5 fragment in the vector. We go back to the pcDNA4/TO vector: as you can see restriction sites are automatically visualized in SnapGene: 1.1 More on restriction analysis in SnapGene.1 Finding restriction sites in the vector.
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