Biochemistry 2280A Lecture Notes - Lecture 4: Polymerase Chain Reaction, Reverse Transcriptase, Autoradiograph

• School: Western University
• Department: Biochemistry
• Course: Biochemistry 2280A
• Professor: Derek Mc Lachlin

Document Summary

What does adding salt (nacl) to the solution do to the tm of the dna: raises the tm. The higher the salt concentration the higher the tm. The higher the sodium ions in the salt shield the repulsive interactions of the negatively charged backbones effectively neutralizing them and raising the tm. Solvents that disrupt hydrogen bonding, interferes with the base pairing and lower the tm. Melting is reversible process is called renaturation, hybridization and annealing. Hybridization will detect complementary nucleic acid sequence s. Hybridizations are often done after transferring the nucleic acid to the membrane. Southern blot: dna to dna on the membrane. Step 1: isolate rna from a group of tissues. Step 2: separate to rna on the basis of size by agrose gel electrophoresis. Step 3: transfer the rna to a nitrocellulose membrane (blotting) The solution is wicked up, which drags the rna from the gel to the paper.