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Journal / Genetics / A Customizable Protocol for String Assembly gRNA…
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Genetics

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JoVE Journal Genetics
A Customizable Protocol for String Assembly gRNA Cloning (STAgR)
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A Customizable Protocol for String Assembly gRNA Cloning (STAgR)

Article DOI: 10.3791/58556-v • 10:00 min • December 26th, 2018
December 26th, 2018 •
1,2, 1,2, 3, 3, 2,4, 1,2
1MCN Junior Research Group, Munich Center for Neurosciences, Ludwig Maximilian Universitat, BioMedical Center, 2Institute of Stem Cell Research, Helmholtz Zentrum, German Research Center for Environmental Health, 3Institute of Developmental Genetics, Helmholtz Zentrum, German Research Center for Environmental Health, 4Physiological Genomics, Ludwig Maximilian Universitat, BioMedical Center

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Summary
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Here, we present string assembly gRNA cloning (STAgR), a method to easily multiplex gRNA vectors for CRISPR/Cas9 approaches. STAgR makes gRNA multiplexing simple, efficient and customizable.

Tags

Customizable Protocol String Assembly GRNA Cloning STAgR Bacterial CRISPR-Cas9 System Guide RNAs Multiplex GRNA Expression Vectors N20 GRNA Sequences Forward Amplification Primers Overhangs Sense GRNA Sequences Reverse Primer Sequences Gibson Assembly High-fidelity Buffer DNTPs Overhang Primers DNA Template String Vector DMSO Water HF Polymerase Thermocycler
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