Recombinant DNA Technology Problem Set

Problem 6: Applications

Tutorial to help answer the question

Restriction endonuclease generated DNA fragments separated by gel electrophoresis and blot transferred onto a membrane filter are probed with a radioactive DNA fragment. This procedure is called:


The Southern Technique

Transferring DNA fragments to nitrocellulose sheets
DNA fragments are transferred to nitrocellulose sheets where they bind.

DNA fragments are denatured and separated by gel electrophoresis.

Fragments are blotted onto a sheet of nitrocellulose and fixed by heating.

Blot is reacted with a radioactive probe of RNA or DNA which binds to complementary DNA.

Autoradiography is used to detect radioactive fragments.

Fixing the fragments of DNA
The denatures fragments of DNA are fixed by baking.

A radioactive probe is added. It can hybridize with a gene sequence in the DNA.

The sheet is rinsed and placed next to X-ray film for autoradiography.

The Biology Project
University of Arizona
Updated: July 15, 1999
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