Hosuk:LabNotes/2014-9-30
Jump to navigation
Jump to search
Tissue Section : fixing test[edit]
- Continued from 2014-09-29
- RCA : start at 2014-09-30, 1pm~ (incubate for overnight)
ppCOL1A1 + Suppressor test[edit]
- Sample : 4 dishes PGP1F, 1st Rolonies made at 2014-07-17
- Check 1st rolonies of all 5 dishes
- S1, S4 --> ppCOL1A1 + suppCOL1A1
- S2, S5 --> ppCOL1A1 only
- Add suppressor 10 times more than ppCOL1A1
File:Fig MIP PGP1F RCA 2014-07-17 1stRolony 3.png
Procedure[edit]
- ppCOL1A1 + supp_COL1A1 : S1, S4
Components | Volume |
5 U/ul Ampligase | 10 |
10X Ampligase Buffer | 10 |
100uM ppCOL1A1 | 2 |
100uM Supp_COL1A1 | 20 |
H2O | 58 |
Total | 100 |
- ppCOL1A1 : S2, S5
Components | Volume |
5 U/ul Ampligase | 10 |
10X Ampligase Buffer | 10 |
100uM ppCOL1A1 | 2 |
100uM Supp_COL1A1 | 0 |
H2O | 78 |
Total | 100 |
- Incubate at 37C for 30min
- Incubate at 60C for 24hrs