mm/dd/yy
- Cas-tone - cell transfections (Gal4EED/luc)
Cas-tone - transfections
- Final H3#_MV11 builds verified on 6/24/15
- Co-transfect H3#_MV11 with pSP-guide RNA plasmid (built by Ben Nyer)
- Use HEK293 Gal4EED/luc cells, no dox (from Rene)
- H340_MV11 + g034_pSPgRNA
- H340_MV11 (no gRNA)
- H360_MV11 + g034_pSPgRNA
- H360_MV11 (no gRNA)
- H380_MV11 + g034_pSPgRNA
- H380_MV11 (no gRNA)
Well |
H3#_MV11 (1.0 μg) |
g034_pSPgRNA (1.0 μg) |
dH2O |
OptiMEM |
PLUS rgt. |
Lipo LTX
|
1 |
5.0 |
1.6 |
13.4 |
570.0 |
2.5 |
7.5
|
2 |
5.0 |
--- |
15.0 |
570.0 |
2.5 |
7.5
|
3 |
5.0 |
1.6 |
13.4 |
570.0 |
2.5 |
7.5
|
4 |
5.0 |
--- |
15.0 |
570.0 |
2.5 |
7.5
|
5 |
5.6 |
1.6 |
12.8 |
570.0 |
2.5 |
7.5
|
6 |
5.6 |
--- |
14.4 |
570.0 |
2.5 |
7.5
|
- Pre-warm ps-free DMEM
- Make 20 μL plasmid/dH2O mixes in sterile 1.5 ml tubes
- Add OptiMEM
- Add PLUS --> 5 min./ RT
- Add Lipo LTX --> 30 min./ RT
- Cells - refresh growth medium w/ ps-free DMEM
- Add 600 μL complexes, drop-wise
- Incubate overnight
|