Ppt1 Knockout cell line (RAW264.7)

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Ppt1 Knockout cell line (RAW264.7)

Ppt1 Knockout cell line (RAW264.7) Order

Catalog#YKO-H834

Price (USD)-

Size 1*10^6

Instruction

Exclusive Red Cotton system(en.rc-crispr.com)was used to design the sgRNA fragment knockout strategy. Two sgRNAs were constructed into CRISPR-U™ plasmid which will be subsequentially transfected into cells by electroporation/virus method. And the single-cell clones were isolated by the limited dilution method and the Monoclone Genotype Validation Kit (Cat: YK-MV-1000) was used for cell lysis and PCR amplification on the single-cell clones, and then the genotype was verified by Sanger sequencing. Upon passing the final validation, the positive clones were expanded and cryopreserved.

Cell Info

Catalog YKO-H834
Cell line Ppt1 Knockout cell line (RAW264.7) Cell Type Mouse Macrophage Cell Line
Morphology monocyte/macrophage, adherent Passage ratio 1:2-1:3
Culture method 90%DMEM+10%FBS
Cryopreservation solution 50%DMEM+40% FBS+10%DMSO
Special Note Cells need to be passaged daily, cannot be digested with trypsin. Cells need to be digested by gently pipetting into single cells.

Gene Information

Official symbol
Ppt1
Gene id
19063
Official full symbol
palmitoyl-protein thioesterase 1
Also known as
9530043G02Rik, AA960502, C77813, CLN1, D4Ertd184e, INCL, PPT
None

KO strategy

Knockout region: Exon 2 (based on transcript Ppt1-201)
Validation: PCR+Sanger Sequencing

Method

Exclusive Red Cotton system(en.rc-crispr.com)was used to design the sgRNA fragment knockout strategy. Two sgRNAs were constructed into CRISPR-U™ plasmid which will be subsequentially transfected into cells by electroporation/virus method. And the single-cell clones were isolated by the limited dilution method and the Monoclone Genotype Validation Kit (Cat: YK-MV-1000) was used for cell lysis and PCR amplification on the single-cell clones, and then the genotype was verified by Sanger sequencing. Upon passing the final validation, the positive clones were expanded and cryopreserved.

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