Talk:20.109(S13):Induce protein and evaluate DNA (Day5)

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(WF O/N Cultures and Gels)
Current revision (12:10, 5 April 2013) (view source)
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Current revision

Contents

Induced pellet example

Examples of induced (left) and control (right) cell pellets
Examples of induced (left) and control (right) cell pellets


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

T/R Red
T/R Red
T/R Orange
T/R Orange
T/R Yellow
T/R Yellow
T/R Green
T/R Green
T/R Blue
T/R Blue
T/R Purple
T/R Purple
T/R Platinum
T/R Platinum


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

W/F Orange
W/F Orange
W/F Yellow
W/F Yellow
W/F Green
W/F Green
W/F Blue
W/F Blue
W/F Pink
W/F Pink
W/F Purple
W/F Purple
W/F Platinum
W/F Platinum


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