Why is it essential that the same restriction enzyme be used to cleave (cut) the DNA of both organisms used to create a transgenic organism?

Question

Oliver Antoine · Accepted Answer

See below...

Restriction enzymes cut at specific sequences so the same restriction enzyme must be used because it will produce fragments with the same complementary sticky ends, making it possible for bonds to form between them.

Sebastian Alvarado · Answer

There are certain compatible restriction sites that can be used together. Xho1 and Sal1 are examples. Their sticky ends match, and so they can be ligated together. But ligation kills both Xho1 and Sal1 site, so you can't use either of these to cut with again (at the ligation locations).

You can also use exonuclease or PCR to make most sticky ends (5or 3 overhangs) into blunt sites - in this way you can ligate into a blunt cut site - but this will generally kill the blunt site, as well.

Jeremiah Allen · Answer

undefined Using the same restriction enzyme ensures compatible DNA ends for successful ligation and gene insertion. Consistent cohesive ends enable the formation of stable recombinant DNA, facilitating the creation of a functional transgenic organism.