09/04/2015
Cells in 23 well plates were well spaced, probably about 70% confluent but proceeded with the transfections anyway. Did g025, g031, g044, and g054. Removed media before and added back media without antibiotics.
| Step 1: Making .5ug/10ul plasmid stocks
|
'
|
'
|
|
|
|
|
1 well |
6.5 wells
|
| ug g034 |
0.5 |
3.25
|
| ul g034 |
|
|
| ul water |
|
|
|
|
65 total ul
|
|
|
|
| Step 2: make DNA + optimem mixes |
|
|
|
1 well |
6.5 x
|
| g034 |
39 |
add 253.5ul optimem
|
|
|
|
| Step 3: add plus reagent to DNA+ opti |
|
|
|
1 well |
6.5 x
|
| g034 |
1 |
add 6.5ul plus reagent
|
|
|
|
| step 4: Make optimem + lipo mastermix |
|
|
|
1 well |
for 24 wells (x 27)
|
| optimem |
47 |
1269
|
| lipo |
3 |
81
|
|
|
|
| Step 5: mix dna mix and lipo mix |
|
|
|
1 well |
6.5 x
|
| g034 |
50 |
add 325 lipo mix
|
|
|
|
| Step 6: add lipo mix to cells |
|
|
| add 100ul of mix to each well |
|
|
Removed media from gal4-eed cells that had been under dox for 96 hours. washed with PBS and replaced with media with puro. washed and replaced media on the puro cells as well. will use these for siRNA transfections.
| 
Project name
Main project page
Previous entry Next entry
