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TYB-SMK-101 | KOD -Plus- Mutagenesis Kit | Cosmo Bio USA available at Bio-Connect | Order online now!

KOD -Plus- Mutagenesis Kit | TYB-SMK-101 | Cosmo Bio USA

KOD -Plus- Mutagenesis Kit

For various mutations, such as substitutions, insertions, and deletion mutations. This kit is an inverse PCR (iPCR)-based site-directed mutagenesis kit using KOD DNA polymerase##<1) 2)>### as a high-fidelity PCR enzyme. This reagent was developed based on a high fidelity and efficient PCR reagent, KOD-Plus- (Code No. KOD-201), which consists of KOD DNA polymerase and anti-KOD DNA polymerase antibodies##<3)>### for Hot Start PCR. This kit enables not only the introduction of point mutations, but also the introduction of large insertions and deletions. The PCR fidelity of KOD-Plus- is greater than Taq DNA polymerase (ca. 80-fold); therefore, unexpected, 2nd-site mutations can be reduced. PCR reactions can be performed using standard PCR primers and do not require phosphorylated primers, because thisprotocol contains a Phosphorylation Step of PCR products. Features: High efficiency (95% maximum) can be obtained. Simple protocol facilitates speedy experiments. Phosphorylated primers are not required. Application: *Site-directed mutagenesis Substitution (e.g. Point mutation, Point mutation library) Deletion (e.g. 1 bp ? several kbp deletion) Insertion (e.g. Introduction of His-tag sequence) Principle: A) Inverse PCR of plasmid DNA, using a mutation primer. B) Plasmid DNA is digested by Dpn I. Note: Dpn I digests methylated DNA, such as plasmid DNA from typical E. coli cell lines (e.g. JM109 and DH5alpha). C) Self-ligation of PCR products is performed by a reaction with T4 polynucleotide kinase and ligase. D) Transformation of E. coli cell lines using self-ligated PCR products.

PP-101 | JBS dNTP-Mutagenesis Kit, Random Mutagenesis by dNTP Analogs | Jena Bioscience products available at Bio-Connect

JBS dNTP-Mutagenesis Kit, Random Mutagen... | PP-101 | Jena Bioscience

JBS dNTP-Mutagenesis Kit, Random Mutagenesis by dNTP Analogs

JBS Mutagenesis Series Within three billion years of evolution, nature has produced a plethora of proteins simply by repeated cycles of random mutagenesis followed by in vivo selection for superior function of the encoded proteins. This example of natural evolution has guided researchers within the last two decades to develop strategies for in vitro permutation of proteins. Among the variety of strategies applied, three major powerful techniques have emerged. Random Mutagenesis by dNTP Analogs This method is based on the incorporation of mutagenic dNTP analogs, such as 8-oxo-dGTP and dPTP, into an amplified DNA fragment by PCR. The mutagenic dNTPs are eliminated by a second PCR step in the presence of the four natural dNTPs only, resulting in a rate of mutagenesis of up to 20%. to JBS dNTP-Mutagenesis Kit #PP-101 Random Mutagenesis by Error-Prone PCR Developed by Caldwell & Joyce (1992) this method introduces mutations in the gene of interest using a PCR reaction under conditions that induce an increased error-rate of the DNA-polymerase. The rate of mutagenesis achieved by error-prone PCR is in the range of 0.6-2.0%. to JBS Error-Prone Kit #PP-102 Random Mutagenesis by DNA Shuffling Developed by Stemmer (1994) DNA shuffling generates libraries by random fragmentation of one gene or a pool of related genes, followed by the reassembly of the fragments in a self-priming PCR reaction. This method allows the recombination of sequences from different, related genes. The overall rate of mutagenesis is approx. 0.7%. to JBS DNA-Shuffling Kit #PP-103 Jena Bioscience now offers all components necessary for each of these techniques ready-to-go in a separate kit, accompanied by a streamlined documentation that maximizes success.

PP-102 | JBS Error-Prone Kit, Random Mutagenesis by Error-Prone PCR | Jena Bioscience products available at Bio-Connect

JBS Error-Prone Kit, Random Mutagenesis ... | PP-102 | Jena Bioscience

JBS Error-Prone Kit, Random Mutagenesis by Error-Prone PCR

PP-103 | JBS DNA-Shuffling Kit, Random Mutagenesis by DNA Shuffling | Jena Bioscience products available at Bio-Connect

JBS DNA-Shuffling Kit, Random Mutagenesi... | PP-103 | Jena Bioscience

JBS DNA-Shuffling Kit, Random Mutagenesis by DNA Shuffling

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