Directed evolution by in vitro compartmentalization

Directed evolution by in vitro compartmentalization

The goal of in vitro compartmentalization (IVC) is to divide a large reaction between many microscopic compartments. This technique was first developed to generate artificial cells for the directed evolution of proteins. Typically, an aqueous solution of genes and an in vitro transcription-translati...

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Veröffentlicht in:Nature methods 2006-07, Vol.3 (7), p.561-570
Hauptverfasser: Miller, Oliver J, Bernath, Kalia, Agresti, Jeremy J, Amitai, Gil, Kelly, Bernard T, Mastrobattista, Enrico, Taly, Valérie, Magdassi, Shlomo, Tawfik, Dan S, Griffiths, Andrew D
Format: Artikel
Sprache:eng
Schlagworte:
Biochemistry, Molecular Biology
Bioinformatics
Biological Microscopy
Biological Techniques
Biomedical and Life Sciences
Biomedical Engineering/Biotechnology
Cell Compartmentation
Cells
Cellular biology
Compartmental analysis (Biology)
Directed Molecular Evolution
Emulsions
Emulsions - chemistry
Flow Cytometry
Gene expression
Genetics
Life Sciences
Microchemistry - methods
Proteomics
protocol
Research methodology
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Beschreibung
Zusammenfassung:The goal of in vitro compartmentalization (IVC) is to divide a large reaction between many microscopic compartments. This technique was first developed to generate artificial cells for the directed evolution of proteins. Typically, an aqueous solution of genes and an in vitro transcription-translation system is stirred (or homogenized) into an oil-surfactant mixture to create a water-in-oil (w/o) emulsion with 1010 aqueous droplets per ml of emulsion. The majority of droplets contain no more than a single gene along with all of the molecular machinery needed to express that gene.
ISSN:1548-7091
1548-7105
DOI:10.1038/nmeth897