QuikChange II-E Site-Directed Mutagenesis Kitįor researchers performing mutagenesis via transformation into electroporation-competent cells. QuikChange II XL Site-Directed Mutagenesis Kitįor long (8 kb – 14kb) or difficult targets. QuikChange II Site-Directed Mutagenesis Kitįor short (4kb – 8kb) targets. Overview of the QuikChange II Site-directed Mutagenesis Method. coli strains is dam methylated and therefore susceptible to Dpn I digestion.) The nicked vector DNA containing the desired mutations is then transformed into competent cells.įigure 1. The Dpn I endonuclease (target sequence: 5´-Gm6ATC-3´) is specific for methylated and hemimethylated DNA and is used to digest the parental DNA template and to select for mutation-containing synthesized DNA.6 (DNA isolated from almost all E. Following temperature cycling, the product is treated with Dpn I. Extension of the oligonucleotide primers generates a mutated plasmid containing staggered nicks. The oligonucleotide primers, each complementary to opposite strands of the vector, are extended during temperature cycling by PfuUltra HF DNA polymerase, without primer displacement. The basic procedure utilizes a supercoiled double-stranded DNA (dsDNA) vector with an insert of interest and two synthetic oligonucleotide primers, both containing the desired mutation (see Figure 1). The QuikChange II site-directed mutagenesis method is performed using PfuUltra high-fidelity (HF) DNA polymerase for mutagenic primer-directed replication of both plasmid strands with the highest fidelity. The QuikChange II site-directed mutagenesis kit is used to make point mutations, replace amino acids, and delete or insert single or multiple adjacent amino acids. Support Services, Agreements & Training.Shorten the time it takes to start seeing the full value of your instrument investment Instrument & equipment deinstallation, transportation, and reinstallationĬrossLab Connect services use laboratory data to improve control and decision-makingĪdvance lab operations with lab-wide services, asset management, relocation Learn essential lab skills and enhance your workflows Instrument/software qualifications, consulting, and data integrity validations Software to manage instrument access, sample processing, inventories, and more ![]() Service Plans, On Demand Repair, Preventive Maintenance, and Service Center Repair Chromatography & Spectroscopy Lab Supplies.
0 Comments
Leave a Reply. |