Overview of Q5 SDM: Site-Directed Mutagenesis using the NEB Q5 SDM Kit скачать в хорошем качестве

Overview of Q5 SDM: Site-Directed Mutagenesis using the NEB Q5 SDM Kit

Q5 SDM: Site-Directed Mutagenesis using the NEB Q5 SDM Kit 1. Perform PCR with 1) a forward primer that incorporates the mutation you want and 2) a reverse primer that abuts the 5' end of the forward primer 2. Add the KLD mixture containing: 1. K: a Kinase to phosphorylate the 5' ends to allow for ligation (since the primers come unphosphorylated) 2. L: A Ligase to stitch the ends together, circularizing the PCR product 3. D: Donl to chew up the unmutated template - Dpni chews up methylated DNA (such as the plasmid, but not the PCR product) Much more on site-directed mutagenesis: https://bit.ly/mutagenesisconstructs  YouTube:     • Mutation mumbo-jumbo: terminology, types, ...       more about all sorts of things: #365DaysOfScience All (with topics listed) 👉 http://bit.ly/2OllAB0 or search blog: http://thebumblingbiochemist.com                                        #scicomm #biochemistry #molecularbiology #biology #sciencelife #science #realtimechem