Lecture 2-3 - Lecture 3 Mutagenesis The Scientific Method...

Info iconThis preview shows pages 1–2. Sign up to view the full content.

View Full Document Right Arrow Icon
Lecture 3 2/21/11 © Nicola Davies 2011 Site Directed vs. Random Mutagenesis Lecture 3 February 21, 2011 2 Site Directed Mutagenesis: 1 Method PCR 2 PCR 1 PCR 3 3 Site Directed Mutagenesis: 1 Method yfg lacZ Amp R Cloning Vector lacZ Ampicillin Resistance MCS PCR 2 PCR 1 PCR 3 + = 4 Site Directed Mutagenesis: 1 Method yfg lacZ Amp R yfg lacZ Amp R Cam R + = Restriction Endonuclease Digest Chloramphenicol Resistance Restriction Endonuclease Digest + Amp R sacB ori * = Amp R sacB ori * yfg Cam R 5 Site Directed Mutagenesis: 1 Method Amp R sacB ori * Homologous Recombination yfg Cam R Bacterial chromosome 6 Enrich for second recombinations by growing in sucrose Two possible sites of homologous recombination Recombine at site A: regenerate WT Recombine at site B: generate a mutant strain 10% sucrose is lethal to strains carrying the sacB gene Patch to find colonies that are Cam R , Amp S A A B B Site Directed Mutagenesis: 1 Method Bacterial chromosome containing mutated copy of yfg
Background image of page 1

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full DocumentRight Arrow Icon
Image of page 2
This is the end of the preview. Sign up to access the rest of the document.

This note was uploaded on 11/17/2011 for the course BIO 456 taught by Professor Smith during the Spring '11 term at Aberystwyth University.

Page1 / 3

Lecture 2-3 - Lecture 3 Mutagenesis The Scientific Method...

This preview shows document pages 1 - 2. Sign up to view the full document.

View Full Document Right Arrow Icon
Ask a homework question - tutors are online