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Daniela Volcan Almeida on Muck Rack

Daniela Volcan Almeida

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Combination of error-prone PCR (epPCR) and Circular Polymerase Extension Cloning (CPEC) for improving the coverage of random mutagenesis libraries - Scientific Reports

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By Natalia Ossa-Hernandez, Luis Fernando Marins, Daniela Volcan Almeida
| Nature Verified
Abstract Random mutagenesis, such as error-prone PCR (epPCR), is a technique capable of generating a wide variety of a single gene. However, epPCR can produce a large number of mutated gene variants, posing a challenge in ligating these mutated PCR products into plasmid vectors. Typically, the primers for mutagenic PCRs incorporate artificial restriction enzyme sites compatible with chosen plasmids. Products are cleaved and ligated to linearized plasmids, then recircularized by DNA ligase.
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