pSpCas9 (PX165)

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	There is no need to perform a negative control golden-gate reaction (without insert) as it will always contain colonies so not a good indicator of cloning success.

Protocol tips
There is no need to perform a negative control golden-gate reaction (without insert) as it will always contain colonies so not a good indicator of cloning success.

Publication protocol

Klf5 gene was engineered as described previously (Ran et al., 2013). Briefly, a plasmid vector containing clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 endonuclease coupled with paired guide RNAs flanking the mouse Klf5 gene was transiently transfected into C2C12 cells. Deletion of targeted loci was confirmed by sequencing. Cells transfected with a plasmid vector carrying CRISPR-Cas9 endonuclease coupled with paired guide RNAs flanking the GFP sequence were used as a control (Abbreviated GFP-targeted control).



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Papers

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Manufacturer protocol

Download the product protocol from Addgene for pSpCas9 (PX165) below.

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Videos

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