3. Screening the vector for insert After generation of the vector, we will transform it into bacteria using electroporation and grow the bacteria on media.
A) note that not all the bacteria will be transformed so what is a quick way we can determine if bacteria have been transformed with the plasmid (hint look at the plasmid)?
B) You digest/cut the plasmid vector with EcoRV, PacI, and MscI. How many bands will be generated using these enzymes and what size will be if you cut an untransformed vector?
C) What about the size of the transformed vector?
D) Draw a gel showing the relative position of the bands generated by your restriction digest from a vector transformed with the insert (from 3c) and one without the insert (make sure to include a marker for comparison)
E) How does the restriction digest indicate you have successfully cloned the
‘gene’ into both MCS regions?

The plasmid is pKannibal: https://www.snapgene.com/plasmids/plant_vectors/pKANNIBAL