pSpCas9(BB)-2A-Puro (PX459)

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

Generation of CRISPR/Cas9-mediated knockout cell lines
L929 cells were transfected with pSpCas9-(BB)2A puro (PX459), a plasmid from Feng Zhang's lab,68 and knockout cells were selected using puromycin at 10 μg/ml for 5 days. Guide oligos used were mouse Sirt2 (5′ primer: CAC CGG CGG AAG TCA GGG ATA CCC G; 3′ primer: AAA CCG GGT ATC CCT GAC TTC CGC C), and mouse Sirt5 (5′ primer: CAC CGA ACT GGG AAA TGA ATC GGC C; 3′ primer: AAA CGG CCG ATT CAT TTC CCA GTT C).

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Reviews

Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing CRISPR Mouse - Deletion L929 SIRT2 using pSpCas9(BB)-2A-Puro (PX459) from Addgene.

Manufacturer protocol

Download the product protocol from Addgene for pSpCas9(BB)-2A-Puro (PX459) below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing CRISPR Mouse - Deletion L929 SIRT2 using pSpCas9(BB)-2A-Puro (PX459) from Addgene. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.