Agilent quikchange manual

The QuikChange HT Protein Engineering System protocol uses a library of custom mutagenic oligonucleotides and custom primers to produce an engineered mutant plasmid library. The mutagenic oligonucleotide library and custom primers must be designed using Agilent’s eArray website before ordering a custom QuikChange HT Protein Engineering System kit. 1 Site Directed Mutagenesis (QuickChange Method) Cornell iGEM Protocol Source: Dylan Webster (Adapted from QuickChange II XL Site-Directed Mutagenesis Kit Protocol). I am a user of QuikChange II XL Site-Directed Mutagenesis Kit (Agilent) and I have performed SDM succesfully several times. I always introduce single nucleotide changes using a pair of mismatched.

b The QuikChange II Site-Directed Mutagenesis Kit (Catalog #) contains enough reagents for 10 total reactions, which includes 5 control reactions. c Thaw the dNTP mix once, prepare single-use aliquots, and store the aliquots at °C. The QuikChange HT Protein Engineering System protocol uses a library of custom mutagenic oligonucleotides and custom primers to produce an engineered mutant plasmid library. The mutagenic oligonucleotide library and custom primers must be designed using Agilent's eArray website before ordering a custom QuikChange HT Protein Engineering System. QuikChange® XL Site-Directed Mutagenesis Kit 5 The QuikChange XL site-directed mutagenesis kit described in this manual is a new version of Stratagene's popular QuikChange site-directed mutagenesis kit. The QuikChange XL kit was created for efficient mutagenesis of large or otherwise difficult-to-mutagenize plasmid templates.

a The QuikChange XL Site-Directed Mutagenesis Kit (Catalog #) contains enough reagents for 30 total reactions, which includes 5 control reactions. b The QuikChange XL Site-Directed Mutagenesis Kit (Catalog #) contains enough reagents for 10 total reactions, which includes 5 control reactions. c See Preparation of Media and Reagents. Note The QuikChange Multi site-directed mutagenesis system is recommended for mutating base pairs. To perform insertions or deletions, use the QuikChange II kit (Stratagene Catalog #) or QuikChange II XL kit (Stratagene Catalog #). Outline of the Three-Step Protocol The three-step QuikChange Multi site-directed mutagenesis method is. The composition of the dNTP mix is proprietary. This reagent has been optimized for the QuikChange II -Esite directed mutagenesis protocols and has been qualified for use in conjunction with the other kit components. Do not substitute with dNTP mixes provided with other Agilent kits.