Information About the DNA to be Cloned

The 2.1kb long insert was excised from the source DNA using NotI. The insert fragment has 2 SphI restriction sites. One of these is located 1.3kb from 1 end of the fragment and the other is located 0.2kb from the other end.

Information About the Vector

The size of the vector is 2.8kb. It has an SphI site located 0.7kb on one side of the NotI site located in the MCS. The vector also has a HindIII site located 1.8kb from the NotI site in the same direction (i.e. on the same side) as the SphI site.

1. Diagram the insert DNA in each of its possible orientations.

2. Diagram the vector.

3. State the total size of the vector plus the insert.

4. Diagram the vector plus insert in both possible orientations of the insert and list the fragment sizes that result from each after the fragment plus insert have been double digested with SphI and HindIII.