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The following cell line-specific conditions are provided as a starting point for transfecting wild-type cells with TrueGuide Synthetic gRNA and TrueCut Cas9 Protein v2 using the Lipofectamine CRISPRMAX Transfection Reagent. Further optimization of the transfection conditions may be necessary for best results.
NOTE: Scroll to the right within the table to see all data.
| Cell type | Source | Media | Cell seeding density/well (× 103) one day before transfection | TrueCut Cas9 Protein v2/gRNA (ng/pmoles) | Lipofectamine Cas9 Plus Reagent/well (μL) | Lipofectamine CRISPRMAX Reagent/well (μL) | ||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Well format | — | — | 96-well | 24-well | 6-well | 96-well | 24-well | 6-well | 96-well | 24-well | 6-well | 96-well | 24-well | 6-well |
| HEK293 | Human embryonic kidney | DMEM | 18 | 90 | 450 | 250/1.5 | 1250/7.5 | 6250/37.5 | 0.5 | 2.5 | 12.5 | 0.4 | 2 | 10 |
| U2OS | Human osteosarcoma | McCoy5A | 10 | 50 | 250 | 250/1.5 | 1250/7.5 | 6250/37.5 | 0.5 | 2.5 | 12.5 | 0.3 | 1.5 | 7.5 |
| A549 | Human epithelial lung carcinoma | DMEM | 10 | 50 | 250 | 250/1.5 | 1250/7.5 | 6250/37.5 | 0.5 | 2.5 | 12.5 | 0.3 | 1.5 | 7.5 |
| THP1 | Human peripheral blood monocyte leukemia | RPMI | 10 | 50 | 250 | 400/2.4 | 2000/12 | 10000/60 | 0.8 | 4 | 20 | 0.3 | 1.5 | 7.5 |
| K562* | Human leukemia bone marrow | RPMI | 10 | 50 | 250 | 250/1.5 | 1250/7.5 | 6250/37.5 | 0.5 | 2.5 | 12.5 | 0.3 | 1.5 | 7.5 |
| iPSC* | Human induced pluripotent stem cells | Essential 8 | 8 | 40 | 200 | 300/2 | 1500/10 | 7500/50 | 0.6 | 3 | 15 | 0.3 | 1.5 | 7.5 |
| HepG2 | Human hepatocellular carcinoma | DMEM | 10 | 50 | 250 | 250/1.5 | 1250/7.5 | 6250/37.5 | 0.5 | 2.5 | 12.5 | 0.3 | 1.5 | 7.5 |
| MDA- MB231 | Human epithelial (breast) adenocarcinoma | DMEM | 10 | 50 | 250 | 250/1.5 | 1250/7.5 | 6250/37.5 | 0.5 | 2.5 | 12.5 | 0.3 | 1.5 | 7.5 |
| N2A | Mouse brain neuroblastoma | DMEM | 10 | 50 | 250 | 250/1.5 | 1250/7.5 | 6250/37.5 | 0.5 | 2.5 | 12.5 | 0.3 | 1.5 | 7.5 |
| *Use the Neon Transfection System for higher editing efficiency. | ||||||||||||||
仅供科研使用,不可用于诊断目的。