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Anonymous
06-04-2004, 09:35 PM
Problem 6 Applications

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


A. Gene cloning
B. The Southern technique
C. The polymerase chain reaction
D. Recombinant DNA
E. Gene mapping




The Southern Technique

Transfering DNA fragments to nitrocellulose sheets

http://www.biology.arizona.edu/molecular_bio/problem_sets/Recombinant_DNA_Technology/graphics/03tb.gif

DNA fragments are transfered 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

http://www.biology.arizona.edu/molecular_bio/problem_sets/Recombinant_DNA_Technology/graphics/03tc.gif

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.



ans:
B. The Southern technique
The Southern technique detects the location of DNA fragments on gel electrophoresis

adam
06-05-2004, 10:25 AM
check the animation at this website:

http://www.dnalc.org/shockwave/southan.html