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A Universal Protocol for Large-scale gRNA Library Production from any DNA Source
JoVE Journal
Genetik
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JoVE Journal Genetik
A Universal Protocol for Large-scale gRNA Library Production from any DNA Source

A Universal Protocol for Large-scale gRNA Library Production from any DNA Source

10,813 Views

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10:32 min

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December 06, 2017

DOI:

10.3791/56264-v

DOI:
10.3791/56264-v

•

10:32 min
December 06, 2017

•

3 Views
,
1MCN Junior Research Group, Munich Center for Neurosciences,Ludwig-Maximilian-Universität, BioMedical Center, 2Epigenetic Engineering, Institute of Stem Cell Research,Helmholtz Zentrum, German Research Center for Environmental Health

Summary

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Methods for generating large-scale gRNA libraries should be simple, efficient and cost-effective. We describe a protocol for the production of gRNA libraries based on enzymatic digestion of target DNA. This method, CORALINA (comprehensive gRNA library generation through controlled nuclease activity) presents an alternative to costly custom oligonucleotide synthesis.

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CRISPRGRNA LibraryGuide RNA LibraryDNA DigestionMicrococcal NucleasePAGEDNA Fragment ExtractionCORALINA MethodDNA Source Agnostic

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Cite this Article

Köferle, A., Stricker, S. H. A Universal Protocol for Large-scale gRNA Library Production from any DNA Source. J. Vis. Exp. (130), e56264, doi:10.3791/56264 (2017).

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