Baculovirus Cre FLP QC 2014 10 09
VLP from sf9 cells.
Purified FLP: Concentrated from 1L to to 200ml (FLP) by ultrafiltration and purified by gel filtration
Original FLP: VLP in sf9 cell culture medium
Purified Cre: Concentrated from 1L to to 200ml (FLP) by ultrafiltration , dialysis and purified by gel filtration, filtered on 10/08/2014
Reporter: 2013 for Flp, 2181 for Cre
Oct. 7 2014 17:00 pm : Transfect 293 cell (70--80%) using lipofectamin 2000
Oct.8 2014 9:00 am: 20%-30% cells turns red , cells are in good condition, add Flp and Cre to reporter cells
|
A1: 25ul Purified FLP |
A2: 50ul Purified FLP |
A3: 75ul Purified FLP |
A4: 150ul Purified FLP |
A5: 75ul Original FLP |
A6: 150ul Original FLP |
|
B1: 25ul Purified Cre |
B2: 50ul Purified Cre |
B3: 75ul Purified Cre |
B4: 100ul Purified Cre |
B5: 150ul Purified Cre |
B6: 200ul Purified Cre |
Oct.9 2014 9:00 am take photos
B4,B5 cell are not vey good
B6 some cells dies, bad
Oct. 7 2014 17:00 pm : Split reporter cells into 24well plate, (2013 from another 24 well plate, 2181 from 10cm plate, transfected on Oct,3)
Oct.8 2014 9:00 am: Flp reporter cells are red ,but in bad condition, only few Cre reporter cells are red
add Flp and Cre to reporter cells
|
A1: 0ul Purified FLP |
A2: 25ul Purified FLP |
A3: 50ul Purified FLP |
A4: 75ul Purified FLP |
A5: 100ul Original FLP |
A6: 150ul Original FLP |
| B1:
200ul Purified FLP |
B2:
50ul Original FLP |
B3:
75ul Original FLP |
B4:
100ul Original FLP |
B5:
100ul Original FLP |
B6:
150ul FIRST FILTERED FLP |
|
C1: 10ul Purified Cre |
C2:
25ul Purified Cre |
C3:
50ul Purified Cre |
C4:
75ul Purified Cre |
C5:
100ul Purified Cre |
C6:
125ul Purified Cre |
| D1:
150ul Purified Cre |
D2:
200ul Purified Cre |
CONTROL |
