BIOL309 Lecture Notes - Lecture 13: Filter Paper, Restriction Enzyme, Plasmid
Document Summary
If screening for lambda libraries, grow on plate that supports the growth of plaques. If screening for plasmid library, grow on plate that supports the growth of colonies. Take a membrane (filter paper) and place it on top, the peel it off gently so some bacteria or some phage particles come off. Save plate, put it in the fridge until experiment is finished. Liberate dna from filter paper so that its available for hybridization to probe, so attach to membrane so it is able to hybridize to incoming dna. Lay membrane on top of some buffer, then bake it. Incubate membrane with solution that contains the probe. Probe: can be dna, rna, or antibody that is going to be able to bind specifically to colonies or phage on membrane. Keep doing until every plaque binds to probe which may take a few weeks. Denature target dna (library, restriction enzyme, mrna samples) Fix denatured dna or rna to membrane.
