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Journal / Genetics / A Universal Protocol for Large-scale gRNA Library …
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Genetics

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

Article DOI: 10.3791/56264-v • 10:32 min • December 6th, 2017
December 6th, 2017 •
1, 1,2
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

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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.

Tags

Universal Protocol Large-scale GRNA Library Production DNA Source Guide RNA Libraries CRISPR-based Screens CORALINA Method Enzymatic Digestion Guide RNA Library Backbone Cost-effective Optimization Micrococcal Nuclease Digestion Reaction Setup Micrococcal Nuclease Buffer Bovine Serum Albumin Target DNA EGTA Sample Buffer Gel Loading Dye Polyacrylamide Gel DNA Ladder TBE Running Buffer
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