Question

How did you use restriction enzymes in your lab and what was accomplished with them?  How was electrophoresis used to elucidate the results of the digest?

Answer 1

Restriction enzymes having several usages, I am going to list them below.

1. Identification of the gene insertion in cloning vector: In cloning experiment vector and PCR or gene of interest is will digest with same restriction enzymes and ligate them. First identification of clone will made by the use of restriction digestion. In this approximately size of the gene and vector is determined.
2. During the plasmid isolation generally good quality of plasmid is supecoiled in nature ant that moves faster in agarose gel electrophoresis hence its band will come below the original size. Some time plasmids show anomalous behaviou in their migration pattern so it becomes difficult to determine their approximate size. By using single restriction site enzyme approximate size of the vector can be determine.
3. In by using common restriction sites one can fuse the two genes.

All the results that come from the restriction sites will be analyze by the agarose gel electrophoresis. Suppose if you have cloned any gene into a new vector then restriction digestion results will be analyzed on gel electrophoresis will give help in the identification of the clone.In that restriction digestion you will get two bands one is insert and one is vector. Here condition that insert and vector size should not be the same.

All the mentioned three examples will analyze in this way.