Anonymous
06-04-2004, 09:20 PM
Problem 3: Southern Technique
The "Southern" technique involves:
A. the detection of RNA fragments on membranes by specific radioactive antibodies.
B. the detection of DNA fragments on membranes by a radioactive DNA probe.
C. the detection of proteins on membranes using a radioactive DNA probe.
D. the detection of proteins on membranes using specific radioactive antibodies.
E. the detection of DNA fragments on membranes by specific radioactive antibodies.
Separating DNA by gel electrophoresis
http://www.biology.arizona.edu/molecular_bio/problem_sets/Recombinant_DNA_Technology/graphics/03t.gif http://www.biology.arizona.edu/molecular_bio/problem_sets/Recombinant_DNA_Technology/graphics/03ta.gif
DNA is taken from individuals A and B
It is digested with restriction endonuclease
Fragments are applied onto an Agarose gel for electrophoresis
DNA has a negative charge, and in an electric field migrates towards a positive electrode.
The rate of migration through a gel is proporational to the size of the fragment.
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 detection of DNA fragments on membranes by a radioactive DNA probe.
The Southern technique detects fragments of DNA using a radioactive DNA probe.
The "Southern" technique involves:
A. the detection of RNA fragments on membranes by specific radioactive antibodies.
B. the detection of DNA fragments on membranes by a radioactive DNA probe.
C. the detection of proteins on membranes using a radioactive DNA probe.
D. the detection of proteins on membranes using specific radioactive antibodies.
E. the detection of DNA fragments on membranes by specific radioactive antibodies.
Separating DNA by gel electrophoresis
http://www.biology.arizona.edu/molecular_bio/problem_sets/Recombinant_DNA_Technology/graphics/03t.gif http://www.biology.arizona.edu/molecular_bio/problem_sets/Recombinant_DNA_Technology/graphics/03ta.gif
DNA is taken from individuals A and B
It is digested with restriction endonuclease
Fragments are applied onto an Agarose gel for electrophoresis
DNA has a negative charge, and in an electric field migrates towards a positive electrode.
The rate of migration through a gel is proporational to the size of the fragment.
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 detection of DNA fragments on membranes by a radioactive DNA probe.
The Southern technique detects fragments of DNA using a radioactive DNA probe.