Mutagenesis Protocols

Creating a Deletion by PCR Splicing Protocol This approach can be used to delete a fragment of any lenth or to introduce point mutations into a DNA seqence.

F2 Mutant Screen - Non "Clonal" Protocol Protocol for F2 mutant screen (non "clonal").

Hydroxylamine Mutagenesis of Plasmid DNA Protocol Protocol for hydroxylamine mutagenesis of plasmid DNA is ideal for random mutagenesis of plasmid DNA which is then used in a plasmid shuffle or screen for ts mutants.

In Vitro Mutagenesis Using Double-stranded DNA Templates: Selection of Mutants with DpnI Protocol for in vitro mutagenesis using double-stranded DNA templates. Two oligonucleotides are used to prime DNA synthesis catalyzed by a high-fidelity thermostable polymerase on a denatured plasmid template. The two oligonucleotides both contain the desired mutation and occupy the same starting and ending positions on opposite strands of the plasmid DNA.

Integrating Extrachromasomal Arrays into the C. elegans Chromosomes Protocol Protocol for integrating extrachromasomal arrays into the C. elegans chromosomes.

Mutagenic PCR Protocol Protocol for mutagenic PCR.

Oligonucleotide-directed Mutagenesis by Elimination of a Unique Restriction Site (USE Mutagenesis) Protocol for oligonucleotide-directed mutagenesis by elimination of a unique restriction site (USE mutagenesis).

Oligonucleotide-directed Mutagenesis of Single-stranded DNA Protocol Protocool for oligonucleotide-directed mutagenesis of single-stranded DNA.

Phosphorylating the 5' Termini of Oligonucleotides Protocol Protocol for phosphorylating the 5' termini of oligonucleotides.

Rapid and Efficient Site-directed Mutagenesis by the Single-tube Megaprimer PCR Method Protocol Protocol for rapid and efficient site-directed mutagenesis by the single-tube megaprimer PCR method.

Rapid PCR Site-Directed Mutagenesis Protocol Protocol for rapid PCR site-directed mutagenesis.

Site Directed Mutagenesis Protocol Protocol for site directed mutagenesis.

Site-specific Mutagenesis by Overlap Extension Protocol Protocol for site-specific mutagenesis by overlap extension.

Systematic Mutagenesis of E.coli K-12 MG1655 ORFs Protocol Protocol for systematic mutagenesis of e. coli K-12 MG 1655.

Transposon Mediated Mutagenesis Protocol Protocol for transposon mediated mutagenesis. Includes: Amplify ORF from MG1655; Transposition reaction; Transformation; Picking; Verify mutation by Culture PCR; Confirm mutations by sequencing; Curing the temperature sensitive pKD46 plasmid.