Biochemistry 2288A Lecture Notes - Lecture 4: Northern Blot, Southern Blot, Reverse Transcriptase

• School: Western University
• Department: Biochemistry
• Course: Biochemistry 2288A
• Professor: Susan Meakin

Document Summary

The + ions in salt shield p backbone (attracted to it) & partially neutralize the repulsion: salt concentration. Cations (+ve charged) shield the repulsive interactions of the phosphate backbone. The higher the salt concentration the higher the tm. Answer: lowers the tm: solvents that disrupt hydrogen bonding such as formamide, urea and formaldehyde disrupt base pairing and lowers the tm. Melting is reversible: process is called renaturation, hybiridzation or annealing). Reversible proess: can go from duplex to single strand then back to duplex. Steps in northern blotting: (strands that aren"t complementary wont anneal/ hybridize) Hybridization experiments are done after transferring the nucleic acis to a membrane. Southern blot: dna to dna on the membrane. Northern blot: dna to rna on the memebrane. Identify which tissues express yfg step 1: isolate rna from a group of tissues step 2: separate the rna on the basis of size by agarose gel electrophoresis.