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Assembling linear DNA templates for in vitro transcription and translation.

Stein, Viktor and Kaltenbach, Miriam and Hollfelder, Florian (2012):
Assembling linear DNA templates for in vitro transcription and translation.
In: Methods in molecular biology (Clifton, N.J.), pp. 67-78, 815, ISSN 1940-6029, [Article]

Abstract

Cell-free expression systems provide straightforward access from genes to the corresponding proteins, involving fewer handling steps than in vivo procedures. A quick procedure to assemble a gene of interest into a linear DNA template together with 3'- and 5'-untranslated regions using a coupled uracil-excision-ligation strategy based on USER Enzyme and T4 DNA ligase. This methodology will be useful for repeated cycles of expression and in vitro selection, in which gene libraries are repeatedly assembled and their products and templates regenerated.

Item Type: Article
Erschienen: 2012
Creators: Stein, Viktor and Kaltenbach, Miriam and Hollfelder, Florian
Title: Assembling linear DNA templates for in vitro transcription and translation.
Language: English
Abstract:

Cell-free expression systems provide straightforward access from genes to the corresponding proteins, involving fewer handling steps than in vivo procedures. A quick procedure to assemble a gene of interest into a linear DNA template together with 3'- and 5'-untranslated regions using a coupled uracil-excision-ligation strategy based on USER Enzyme and T4 DNA ligase. This methodology will be useful for repeated cycles of expression and in vitro selection, in which gene libraries are repeatedly assembled and their products and templates regenerated.

Journal or Publication Title: Methods in molecular biology (Clifton, N.J.)
Volume: 815
Divisions: 10 Department of Biology
10 Department of Biology > Protein Engineering of Ion Conducting Nanopores
Date Deposited: 14 Nov 2016 11:35
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