BIOL 301 Lecture Notes - Lecture 4: Amplicon, In Situ Hybridization, Ubiquitin
23 Apr 2014
School
Department
Course
Professor
Document Summary
gene expression analysis transcript level: datamining, gross tissue-based techniques, rna detection directly in tissues. It just means you have something disrupting lacz sequence. Thus, you need to actually confirm that those colonies actually has vector and insert. There are few ways to do this: colony pcr: take your pipet tip and touch white colony and throw it into pcr reaction mix. You design 1 primer that will recog insert sequence and the other that recog vector (see hand-drawn pic on the side). Once have a clone, we have to ensure that the clone has the right insert. selectable marker & insertion marker only tell you that you have a plasmid with an inserted piece of dna . If you happen to touch colony next to the colony you picked, you can actually have two diff colonies. 2: restriction enzyme digests - the one we will use.
