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 line||MTHFD1 Knockout cell line(HEK293)||Cell Type||Human Embryonic Kidney Cell Line|
|Morphology||Epithelial-like, adherent||Passage ratio||1:4~1:6|
|Culture method||90%DMEM+10% FBS|
|Cryopreservation solution||50%DMEM+40% FBS+10%DMSO|
has low passages, high activity and good cell condition, STR Authentication reports availableview detail >
EZ-cryo™ Cell Freezing Medium, Ready-to-use, serum-free, protein-free, up to 90% viability rate upon thawingview detail >
For gene-editing mice. Extraction - free, 15 mins for PCR reaction, high-throughput operation on 96-well plate.view detail >