Question

A linear 4.0 kilobase pair (kbp) sequence of DNA was treated the restriction enzymes HindIII and BamHI individually as well as in combination. The reaction products of the three reactions were then run on an agarose gel to resolve the restriction fragments based on size. Using the results of the agarose gel below, construct a restriction map for the sample of DNA. 1 Restriction Enzyme HindIII BamHI HindIII + BamHI Fragments (kbp) 2.8; 1.2 1.8; 1.3; 0.9 1.8; 1.0; 0.9; 0.3

Answer 1

Ans. Steps for generating restriction map of a linear DNA-

I. Draw the restriction fragments of the combined restriction (HindIII + BamHI).

II. Draw the restriction fragments of individual restrictions (HindIII alone, and BamHI alone) up and down the combined restriction.

III. Overlap the restriction sites in three restriction drawings (overlapping shown as dotted lined).

IV. Generate the restriction map from overlapping as shown in figure.