NF1 Knockout cell line (U-87 MG)

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NF1 Knockout cell line (U-87 MG)

NF1 Knockout cell line (U-87 MG) Order

Catalog#YKO-H833

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-H833
Cell line NF1 Knockout cell line (U-87 MG) Cell Type Human Osteosarcoma Cell Line
Morphology Epithelial-like, adherent Passage ratio 1:3~1:5
Culture method 90% MEM+10%FBS
Cryopreservation solution 50%MEM+40%FBS+10%DMSO
Special Note

STR Validation

Gene Information

Official symbol
NF1
Gene id
4763
Official full symbol
neurofibromin 1
Also known as
NFNS, VRNF, WSS
This gene product appears to function as a negative regulator of the ras signal transduction pathway. Mutations in this gene have been linked to neurofibromatosis type 1, juvenile myelomonocytic leukemia and Watson syndrome. The mRNA for this gene is subject to RNA editing (CGA>UGA->Arg1306Term) resulting in premature translation termination. Alternatively spliced transcript variants encoding different isoforms have also been described for this gene.

KO strategy

Knockout region: Exon 12 (based on transcript NF1-202)
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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