JBS DNA-Shuffling Kit, Random Mutagenesis by DNA Shuffling

Jena Bioscience

JBS DNA-Shuffling Kit, Random Mutagenesis by DNA Shuffling

5

Research Use Only

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.

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