pSpCas9(BB)-2A-GFP (PX458)

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

Transfection and cell sorting of MKO-PC12 cells
PC12 cells were cultured in six-well dishes to 70–80% confluence. Cells were transfected with 1 µg sequence-verified pSpCas9-Munc18-1 (sgRNA)-2A-GFP using Lipofectamine 2000 (Invitrogen). 48 h after transfection, cells were pelleted in DMEM and sorted in 96-well plates using a FACSAris II cell sorter (BD). Single cells from GFP-expressing cells (high expression population) were expanded to obtain individual clones.

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Papers

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Manufacturer protocol

Download the product protocol from Addgene for pSpCas9(BB)-2A-GFP (PX458) below.

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