GeneArt™ CRISPR Nuclease Vector with OFP Reporter Kit

CRISPR Human - Deletion TRIM25

Experiment

CRISPR Human - Deletion TRIM25

Product

GeneArt™ CRISPR Nuclease Vector with OFP Reporter Kit from Thermo Fisher Scientific

Manufacturer

Thermo Fisher Scientific

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Protocol tips

Upstream tips	Protocol tips	Downstream tips
	The presence of PEG and glycerol (supplied by the Ligation Buffer and the T4 DNA Ligase) will make the reaction mixture viscous. Be sure to mix the reaction thoroughly by pipetting up and down. Do not vortex.

Protocol tips
The presence of PEG and glycerol (supplied by the Ligation Buffer and the T4 DNA Ligase) will make the reaction mixture viscous. Be sure to mix the reaction thoroughly by pipetting up and down. Do not vortex.

Publication protocol

Generation of TRIM25-knockout HeLa cells
We used the GeneArt® CRISPR Nuclease (OFP Reporter) Vector Kit (Thermo Fisher Scientific) according to the manufacturer's instructions to generate TRIM25 CRISPR nuclease constructs, which were transfected into HeLa cells with Effectene transfection reagent (Qiagen). TRIM25-knockout HeLa cells clones were identified by immunoblot analysis.

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Reviews

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Discussion

4 years ago

Author: Milena Alexeyeva Russian Federation

DNA insert using CRISPR

I would like to excise a large strand of DNA and insert a new one using CRISPR. My problem is that my strand will be a little over 1kb and I am not sure if this is going to be a limiting factor. Also, how long should the homology arms be for a region of this size?

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Papers

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for GeneArt™ CRISPR Nuclease Vector with OFP Reporter Kit below.

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Videos

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