01-05-2007, 06:40 PM
I. Cleavage of genomic DNA by treatment with a specific endonuclease/fragments separated according to size by agoarose gel electrophoresis.
1. So first we have genomic DNA, then 2.DNA fragments with a process of restriction digestion and the 3.digested DNA apears a s acontinous smear on the agaros egel-large fragments in th etop, small fragments on the bottom
II.Denaruration of the DNA Fragments by treatment of alkali
1.gel place don a buffer -satureted filter paper + cover with a sheet of nitrocelulose +overlaid with dry filter paper
III.Immobilization on nitrocellulose
1.buffer carry single straneded DNA
2.filter paper draw buffer out
3.DNA binds to nitocellulose like a happy couple
III.Detection of a specific DNA fragment by hybridization
1.filter is dry and incubated with radioactive DNA probe solution that is comlementary to the sequance of the DNA of intrest.
2.Visualization of hybridazation is achievd by autoradiography.
Southern Blotting is used to detect DNA Fragments in a agorose gel by hybrydization with a specific radiolabelad ologonucleotid eprobe.
Any questions ?
1. So first we have genomic DNA, then 2.DNA fragments with a process of restriction digestion and the 3.digested DNA apears a s acontinous smear on the agaros egel-large fragments in th etop, small fragments on the bottom
II.Denaruration of the DNA Fragments by treatment of alkali
1.gel place don a buffer -satureted filter paper + cover with a sheet of nitrocelulose +overlaid with dry filter paper
III.Immobilization on nitrocellulose
1.buffer carry single straneded DNA
2.filter paper draw buffer out
3.DNA binds to nitocellulose like a happy couple
III.Detection of a specific DNA fragment by hybridization
1.filter is dry and incubated with radioactive DNA probe solution that is comlementary to the sequance of the DNA of intrest.
2.Visualization of hybridazation is achievd by autoradiography.
Southern Blotting is used to detect DNA Fragments in a agorose gel by hybrydization with a specific radiolabelad ologonucleotid eprobe.
Any questions ?