Talk:20.109(S13):Induce protein and evaluate DNA (Day5)
Induced pellet example
TR O/N Cultures and Gels
Below you should list the OD600 values of the induced cultures (stock value), as well as your pellet observations. (Is the color dull or bright? More yellow or more green?) The teaching faculty will fill in this information for the cultures needing to be grown overnight.
| Group | OD WT | OD D24H/E67K/T79P/M124S | OD Chosen mutant | Pellets yellow? |
|---|---|---|---|---|
| Example | 0.95 | 1.2 (D24H) | 0.82 (G20P) | Y, bright ylw-grn |
| Red | 0.22 | 0.51 (D24H) | 0.55 | grn, grn, ylw (+hint of grn) |
| Orange | 0.36 | 0.36 (D24H) | 0.38 | grn, grn, ylw (+hint of grn) |
| Yellow | 0.39 | 2.12 (T79P) | 3.36 (K77E2--not the chosen mutant) extra is 0.17 |
bright yellow; ylw (+hint of grn) |
| Green | 0.34 | 1.59 (T79P) | 1.16 (M71R) | Bright Yellow |
| Blue | 0.035 | 0.036 (M124S) | 0.088 (M71S) | Y, bright ylw |
| Purple | 0.30 | 0.58 (M124S) | 0.32 | ylw-grn, ylw-grn, grn |
| Platinum | 0.52 | 0.75 (M124S) | 0.38 | ylw-grn, ylw-grn, grn |
Gels
WF O/N Cultures and Gels
Below you should list the OD600 values of the induced cultures (stock value), as well as your pellet observations. (Is the color dull or bright? More yellow or more green?) The teaching faculty will fill in this information for the cultures needing to be grown overnight.
Regarding color, please check your pellets when you receive them at the beginning of D6. The small ones were all yellow or green, while some large pellets appeared to be a dull tan color. Upon inspection by fluorescent microscopy (thanks to Shannon's quick thinking), it was clear that a fluorescent protein was indeed present, but masked by the amount of cells and debris.
| Group | OD WT | OD D24H/E67K/T79P/M124S | OD mutant option 1 | OD mutant option 2 | Pellets yellow? |
|---|---|---|---|---|---|
| Example | 0.95 | 1.2 (D24H) | 0.82 (G20P) | N/A | Y, bright ylw-grn |
| Red | 4.94 | 0.40, 3x (D24H) | 0.59, 3x (Blu #1) | N/A | |
| Orange | 4.76 | 0.61 (D24H) | 4.65 | 4.64 | |
| Yellow | 4.86 | 4.47 (M124S) | 0.54, 3x | 0.81, 3x | |
| Green | 4.57 | 0.43, 3x (T79P) | 0.40 | 4.82 | |
| Blue | 4.95 | 4.59 (D24H) | 3.7 (#2) | N/A | |
| Pink | 5.19 | 0.78, 3x (D24H) | 0.64, 3x | 0.50? | |
| Purple | 5.14 | 1.89(M124S) | 0.67, 3x | 4.93 | |
| Platinum | .1 | .091 (E67K) | .068 |
Gels
