Site-Directed Mutagenesis Kit

Description

The Site-Directed Mutagenesis Kit can induce mutagenesis at the specific point of sequence that cloned on plasmid DNA. It guarantees 100% of efficiency in theory. Also it is very convenient and simple because it takes just two steps for all experimental procedures. The Site-Directed Mutagenesis Kit does not necessary using M13 vector and methylation step. Indeed, the Kit can induce mutation of nucleotide, re-mutation to wild type, mutation of codon and insertion even deletion. As the Kit has these characteristics, it is applicable to analysis for genomic/proteomic function. Also as inducingmutagenesis of specific gene, it can be used for protein engineering like protein development or improving productivity.

When you use this the Kit, you can have mutated clone as doing simple steps. (Design primer with own protocol, use the Enzyme for 15~18 cycles of PCR. Proceed transformation step after the Mutazyme treatment for mutated clone selection) In this theory, clones on LB agar plate are mutated around 100% and after sequencing, you can proceed to the next step.

Characteristics

▪ Without special skill, easy to use.
▪ Can induce mutagenesis within 2~3 days.
▪ Use only two enzymes: Muta-direct™ Enzyme and Mutazyme™ Enzyme.
▪ Can use for various experiment: Point mutation, Deletion, Insertion and etc.
▪ 100% of mutation efficiency.
▪ Reasonable price.
▪ Technical assist by research agents.

Storage and Stability

All component should be stored at -20℃. The reaction buffer and dNTP mixture have been optimized for the Muta-direct™ protocols. Do not substitute with buffers or dNTP mixture provided with other kits.

Kit Contents

For research use. The Muta-direct™ kit contains sufficient reagents to perform approximately15×50μl mutagenesis reactions. The kits contain enough control template and primer mix for 5 control reac-tion, and enough reagents for 15 reactions total (control and experimental reactions combined).

Muta-direct™ Control Reaction

Control plasmid, contained in Muta-direct™, is pUC18 that informs us whether the experiment success or not. pUC18 plasmid has lacZ gene, so we can confirm the result as induce termination codon at lacZ gene by using Control Primer Mix (provided). In case of success, there must be all white colonies on LB plate. As change from serine (TCG) to stop codon (TAG) in pUC18, lacZ gene can be blocked.

If user handles the mutation procedure for the first time, he can know about result as proceed of the control reaction step.

Muta-direct Protocol