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||ZEB1 Knockout cell line(THP-1)||Cell Type||Human Ovarian Granulosa Cell Line|
|Morphology||Monocyte, suspension||Passage ratio||Control the cell density at 2x10^5~4.0x10^5 cells/ml during passaging, and carry out the passage when the cells grow to 8x10^5~1.0x10^6 cells/ml.|
|Special Note||The cell culture density needs to be strictly controlled. The cell viability after thawing is relatively low, it is recommended to use Australia-sourced FBS for thawing or increase the FBS ratio to 20%. When the cells grow normally and can be passaged after 2 passages, the cells can be cultured using the normal culture method.|
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 >