Question

How to construct or figure out a restriction digest map

Answer 1

Restriction map is used to find out the sequence of long DNA molecule by cutting it into numerous fragments using restriction enzymes. The steps are as follows:

1.      Take a large DNA suppose 1500bp.

2.      DNA needs to cut with 2-3 restriction enzymes in separate tubes generating fragments of different sizes.

3.      Each garment will be separated by gel electrophoresis and should be collected in individual tube for mapping.

4.      Each DNA fragment will be mapped by any of the available technique (Sanger sequencing method etc.)

5.      Align all the sequences of different fragments by using common sequences in DNA fragments to generate whole sequence (See image attached).