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

Protocol tips

Upstream tips	Protocol tips	Downstream tips
To access a plasmid, keep the plate on dry ice to prevent thawing. Using a sterile pipette tip (20uL or 200uL one), puncture the seal above an individual well and spread a portion of the glycerol stock onto an agar plate.

Upstream tips
To access a plasmid, keep the plate on dry ice to prevent thawing. Using a sterile pipette tip (20uL or 200uL one), puncture the seal above an individual well and spread a portion of the glycerol stock onto an agar plate.

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

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Manufacturer protocol

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

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Videos

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