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JoVE Journal Biology
En protokol for Funktionel vurdering af Whole-Protein Saturation mutagenese Biblioteker hjælp High-Throughput Sequencing
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En protokol for Funktionel vurdering af Whole-Protein Saturation mutagenese Biblioteker hjælp High-Throughput Sequencing

Article DOI: 10.3791/54119-v • 11:36 min • July 3rd, 2016
July 3rd, 2016 •
1, 1, 1, 1
1Green Center for Systems Biology, University of Texas Southwestern Medical Center

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Vi præsenterer en protokol for den funktionelle vurdering af omfattende single-site mætning mutagenese biblioteker af proteiner udnytte high-throughput sekventering. Vigtigere er det, denne fremgangsmåde bruger ortogonale primerpar at multiplekse bibliotekskonstruktion og sekventering. Repræsentative resultater ved hjælp af TEM-1 β-lactamase valgt på en klinisk relevant dosering af ampicillin leveres.

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Molecular Biology mutagenese mætning mutagenese næste generation sekventering high throughput sekventering TEM-1 beta-lactamase antibiotikaresistens ortogonale primere
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