BIO370Y5 Lecture Notes - Lecture 4: Bamhi, Hindiii, Plasmid
Document Summary
Plasmids from part a, control samples of pamp and pkan are incubated with bamhi and hindiii. Uncut, control and mini-prep dna and bsteii and the rest of the ligation will be electrophoresed. Will tell us the sizes, structure and number of plasmids in each transformed strain. Over incubation may cause dnases in mini-prep to degrade plasmid dna. Miniprep lanes contain this: a background smear of degraded and partially digested chromosomal dna, plasmid. Dna and rna: undissolved or high-molecular weight dna trapped at the front edge of the well, a cloud of low weight rna at the 100-200 bp area, presence of high-molecular weight, uncut plasmid of digested mini-prep dna. We used bamhi to digest the marker rather than bsteii. Some facts to remember about m1 and m2 reproduction: every replicating plasmid has an origin of replication. It creates inverted repeats (= a nucleotide sequence downstream of the plasmid) and causes a hairpin to occur.
