Talk:20.109(F14): Mod 2 Day 7 Readout of Protein: Difference between revisions

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==Strains #s and Mutations==
*466 = wt
*468 = Kinase dead
*486 = T541R
*487 = T541Y
*488 = T541P
*489 = T541R
*491 = T541L
*492 = T541silent change
==T/R b-gal data==
==T/R b-gal data==
Be sure to add the mutant strain # when you fill in this chart!
Be sure to add the mutant strain # when you fill in this chart!
Here are the Miller Unit values for WT and KD data:
WT dark = 728.93, 804.09
WT light = 449.42, 420.19
KD dark = 77.67, 77.67
KD light = 55.23, 57.14


{| border="1"
{| border="1"
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|Units
|Units
|-
|-
|<font color = red>Red</font color>
|<font color = red>Cherries</font color>
|WT
|WT
|
|
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|-
|-
|
|
|mut1
|mut1: 487
|
|.27 light, .25 dark
|
|2.7 light, 2.5 dark
|  
|18.83 light, 19 dark
|
|.552 light, .587 dark
|
|.0045 light, .0045 dark
|
|1070 light, 1218 dark
|-
|-
|
|
|mut2
|mut2: 491
|
|.26 light, .24 dark
|
|2.6 light, 2.4 dark
|  
|19.83 light, 19.83 dark
|
|.670 light, .608 dark
|
|.0075 light, .006 dark
|
|1273 light, 1255 dark
|-
|-
|}
|}
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|Units
|Units
|-
|-
|<font color = orange>Orange</font color>
|<font color = orange>Tiger</font color>
|WT
|WT
|
|
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|-
|-
|
|
|mut1
|mut1: 488
|
|Light: 0.29, Dark: 0.24,
|
|Light: 2.9, Dark: 2.4
|  
|Light: 15.33, Dark: 10.33,
|
|Light: 0.4285, Dark: 0.3536,
|
|Light: 0.0055, Dark: 0.006,
|
|Light: 942.2, Dark: 1393.6,
|-
|-
|
|
|mut2
|mut2: 491
|
|Light: 0.26, Dark: 0.3
|
|Light: 2.6, Dark: 3.0
|  
|Light: 10, Dark: 15.17
|
|Light: 0.4285, Dark: 0.586
|
|Light: 0.005, Dark: 0.008
|
|Light: 1614.4, Dark: 1256.9
|-
|-
|}
|}
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|Units
|Units
|-
|-
|<font color = yellow>Yellow</font color>
|<font color = gold>Yellow</font color>
|WT
|WT
|
|
Line 126: Line 146:
|-
|-
|
|
|mut1
|mut1: 488
|
|L: 0.22, D: 0.205
|
|L: 2.2, D: 2.05
|  
|29.83 (for all)
|
|L1: 0.723; L2: 0.637; D1: 0.873; D2: 0.899
|
|L1: -0.001; L2: 0.000; D1: 0.000; D2: 0.001
|
|L1: 1104.36; L2: 970.65; D1: 1427.60; D2: 1467.26
|-
|-
|
|
|mut2
|mut2: 491
|
|L: 0.24, D: 0.3
|
|L: 2.4, D: 3.0
|  
|29.83 (for all)
|
|L1: 1.12; L2: 1.137; D1: 1.041; D2: 0.997
|
|L1: 0.002; L2: 0.002; D1: 0.001; D2: 0.006
|
|L1: 1559.53; L2: 1583.28; D1: 1161.30; D2: 1102.36
|-
|-
|}
|}
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|-
|-
|
|
|mut1
|mut1 (486)
|
|L= 0.28, D= 0.26
|
|L=2.8, D=2.6
|  
|L1= 3.32, L2= 3.32, D1= 2.83, D2= 3.13
|
|L1= 0.688, L2= 0.632, D1= 0.604, D2= 0.485
|
|L1= 0.054, L2= 0.042, D1= 0.061, D2= 0.039
|
|L1= 6390.88, L2= 6014.00, D1= 6750.00, D2= 5115.59
|-
|-
|
|
|mut2
|mut2 (487)
|
|L= 0.24, D= 0.29
|
|L= 2.4, D=2.9
|  
|L1= 3.28, L2= 3.27, D1= 3.33, D2= 3.32
|
|L1= 0.643, L2= 0.560, D1= 0.526, D2= 0.459
|
|L1= 0.009, L2= 0.012, D1= 0.008, D2= 0.007
|
|L1= 7960.03, L2= 6875.00, D1= 5296.55, D2= 4644.78
|-
|-
|}
|}
Line 220: Line 240:
|-
|-
|
|
|mut1
|mut1 (486)
|
|Light1 = 2.4, Dark1 = 2.9
|
Light2 = 2.4, Dark2 = 2.9
|  
|Light1 = 0.24, Dark1 = 0.29
|
Light2 = 0.24, Dark2 = 0.29
|
|17 (across all samples)
|
|Light1 = 0.590, Dark1 = 0.598
Light2 = 0.572, Dark2 = 0.615
|Light1 = 0.007, Dark1 = 0.008
Light2 = 0.009, Dark2 = 0.009
|Light1 = 1419, Dark1 = 1184
Light2 = 1363, Dark2 = 1215
|-
|-
|
|
|mut2
|mut2 (487)
|
|Light1 = 2.8, Dark1 = 2.6
|
Light2 = 2.8, Dark2 = 2.6
|  
|Light1 = 0.28, Dark1 = 0.26
|
Light2 = 0.28, Dark2 = 0.26
|
|17 (across all samples)
|
|Light1 = 0.650, Dark1 = 0.680
Light2 = 0.641, Dark2 = 0.679
|Light1 = 0.006, Dark1 = 0.007
Light2 = 0.006, Dark2 = 0.007
|Light1 = 1343, Dark1 = 1510
Light2 = 1324, Dark2 = 1508
|-
|-
|}
|}
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|-
|-
|
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|mut1
|mut1-487
|
|L1&2-0.28, D1&2-0.26
|
|L1&2-2.8, D1&2-2.6
|  
|L1&2-18.33, D1&2-17.67
|
|L1-0.609, L2-0.616, D1-0.632, D2-0.621
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|L1-0.007, L2-0.003, D1-0.006, D2-0.006
|
|L1-1162.7, L2-1190.0, D1-1352.8, D2-1328.8
|-
|-
|
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|mut2
|mut2-488
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|L1&2-0.22, D1&2-0.25
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|L1&2-2.2, D1&2-2.5
|  
|L1&2-25.17, D1&2-25.17
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|L1-0.605, L2-0.607, D1-0.743, D2-0.756
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|L1-0.01, L2-0.005, D1-0.006, D2-0.006
|
|L1-1061.0, L2-1080.4, D1-1164.1, D2-1184.7
|-
|-
|}
|}
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|-
|-
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|mut1
|mut1 (486)
|
|L: 0.18; D: 0.22
|
|L: 1.8; D: 2.2
|  
|35 (for all)
|
|L1: 0.819; L2: 0.818; D1: 0.831; D2: 0.886;
|
|L1: 0.003; L2: 0.002; D1: 0.001; D2: 0.004
|
|L1:1292.67; L2: 1292.86; D1: 1076.95; D2: 1141.56
|-
|-
|
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|mut2
|mut2 (488)
|
|L: 0.22; D: 0.205
|
|L: 2.2; D: 2.05
|  
|35 (for all)
|
|L1: 0.779; L2: 0.78; D1: 1.08; D2: 1.004
|
|L1: 0.002; L2: 0.006; D1: 0.004; D2: 0.004
|
|L1: 1007.14; L2: 999.35; D1: 1495.47; D2: 1389.55
|-
|-
|}
|}
[[Image:Purple_M2D7_blot.jpg]]
|TR Purple Group Western Blot; Lane 1: H6 Envz, Lane 2: Wild Type, Lane 3: T541T Lane 4: 488- T541T


==W/F b-gal data==
==W/F b-gal data==
Line 349: Line 382:
|WT
|WT
|
|
|
|D-1:.269, 2:.269, 3:.269, L-1:.273,2:.273,3:.273
|  
|D-1:21.5, 2:21.5, 3:21.5, L-1:21.5,2:21.5,3:21.5
|
|D-1:.665, 2:.658, 3:.666, L-1:.305,2:.299,3:.286
|
|D-1:.011, 2:.005, 3:.006, L-1:.013,2:.01,3:.009
|
|D-1:1116.538, 2:1122.59, 3:1133.397, L-1:480.8757,2:479.5979,3:460.431
|-
|-
|
|
|KD
|KD
|
|
|
|D-1:.226,2:.226,3:.226, L-1:.254,2:.254,3:.254
|  
|D-1:21.5,2:21.5,3:21.5, L-1:21.5,2:21.5,3:21.5
|
|D-1:.036,2:.035,3:.037, L-1:.04,2:.04,3:.043
|
|D-1:.005,2:.004,3:.006, L-1:.005,2:.005,3:.007
|
|D-1:56.0815,2:57.62503,3:54.53797, L-1:56.56621,2:57.22395,3:56.30837
|-
|-
|
|
|mut1
|mut1 (487, T541Y)
|
|
|
|
|
|
|
|D-1:.23, 2:.23, 3:.23, L-1:.261,2:.261,3:.261
|D-1:21.5, 2:21.5, 3:21.5, L-1:21.5,2:21.5,3:21.5 
|D-1:.717, 2:.771, 3:.788, L-1:.726,2:.772,3:.783
|D-1:.01, 2:.01, 3:..011, L-1:..012,2:.012,3:.011
|D-1:1414.56, 2:1523.761, 3:1554.601, L-1:.726,2:.772,3:.783
|-
|-
|
|
|mut2
|mut2 (488, T541P)
|
|
|
|
|
|
|
|D-1:.226, 2:.226, 3:.226, L-1:.252,2:.252,3:.252
|D-1:21.5, 2:21.5, 3:21.5, L-1:21.5,2:21.5,3:21.5 
|D-1:.645, 2:.66, 3:.673, L-1:.671,2:.66,3:.64
|D-1:.008, 2:.01, 3:.01, L-1:.01,2:.008,3:.011
|D-1:.1298.621, 2:1322.289, 3:1349.043, L-1:1206.165,2:1192.322,3:1145.718
|-
|-
|}
|}
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|-
|-
|<font color = blue>Blue</font color>
|<font color = blue>Blue</font color>
|WT
|WT NB466
|
|0.263, 0.255
|
|(*light, dark*)
|  
|17.99, 18.00
|
|0.221, 0.319
|
|0.00433, 0.004
|
|450.8, 1162.2 (stdev 11.7, 37.4)
|-
|-
|
|
|KD
|KD NB468
|
|0.249, 0.247
|
|(*light, dark*)
|  
|17.99, 18.00
|
|0.019, 0.026
|
|0.0035, 0.00367
|
|28.6, 53.8 (stdev 1.96, 2.72)
|-
|-
|
|
|mut1
|mut1 NB486 (T541R)
|
|0.199, 0.199
|
|(*light, dark*)
|  
|7.67, 7.67
|
|0.266, 0.267
|
|0.01, 0.007
|
|1622.8, 1667.6 (stdev 128.6, 15.7)
|-
|-
|
|
|mut2
|mut2 NB488 (T541P)
|
|0.257, 0.223
|
|(*light, dark*)
|  
|8.67, 7.67
|
|0.27, 0.253
|
|0.005, 0.00467
|
|1174.4, 1432.1 (stdev 20.5, 8.99)
|-
|-
|}
|}
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|-
|-
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|}
WF Blue Group Western Blot
(Mutant 1: no mutation, Mutant 2: T541S)
[[Image: BlueWFWestern.jpg|left|400x600px]]

Latest revision as of 20:41, 10 November 2014

Strains #s and Mutations

  • 466 = wt
  • 468 = Kinase dead
  • 486 = T541R
  • 487 = T541Y
  • 488 = T541P
  • 489 = T541R
  • 491 = T541L
  • 492 = T541silent change

T/R b-gal data

Be sure to add the mutant strain # when you fill in this chart!

Here are the Miller Unit values for WT and KD data:

WT dark = 728.93, 804.09

WT light = 449.42, 420.19

KD dark = 77.67, 77.67

KD light = 55.23, 57.14

Team Strain OD600 (1:10) OD600 Elapsed time (minutes) Abs420 (list both values) OD550 (list both values) Units
Cherries WT
KD
mut1: 487 .27 light, .25 dark 2.7 light, 2.5 dark 18.83 light, 19 dark .552 light, .587 dark .0045 light, .0045 dark 1070 light, 1218 dark
mut2: 491 .26 light, .24 dark 2.6 light, 2.4 dark 19.83 light, 19.83 dark .670 light, .608 dark .0075 light, .006 dark 1273 light, 1255 dark
Team Strain OD600 (1:10) OD600 Elapsed time (minutes) Abs420 (list both values) OD550 (list both values) Units
Tiger WT
KD
mut1: 488 Light: 0.29, Dark: 0.24, Light: 2.9, Dark: 2.4 Light: 15.33, Dark: 10.33, Light: 0.4285, Dark: 0.3536, Light: 0.0055, Dark: 0.006, Light: 942.2, Dark: 1393.6,
mut2: 491 Light: 0.26, Dark: 0.3 Light: 2.6, Dark: 3.0 Light: 10, Dark: 15.17 Light: 0.4285, Dark: 0.586 Light: 0.005, Dark: 0.008 Light: 1614.4, Dark: 1256.9
Team Strain OD600 (1:10) OD600 Elapsed time (minutes) Abs420 (list both values) OD550 (list both values) Units
Yellow WT
KD
mut1: 488 L: 0.22, D: 0.205 L: 2.2, D: 2.05 29.83 (for all) L1: 0.723; L2: 0.637; D1: 0.873; D2: 0.899 L1: -0.001; L2: 0.000; D1: 0.000; D2: 0.001 L1: 1104.36; L2: 970.65; D1: 1427.60; D2: 1467.26
mut2: 491 L: 0.24, D: 0.3 L: 2.4, D: 3.0 29.83 (for all) L1: 1.12; L2: 1.137; D1: 1.041; D2: 0.997 L1: 0.002; L2: 0.002; D1: 0.001; D2: 0.006 L1: 1559.53; L2: 1583.28; D1: 1161.30; D2: 1102.36
Team Strain OD600 (1:10) OD600 Elapsed time (minutes) Abs420 (list both values) OD550 (list both values) Units
Green WT
KD
mut1 (486) L= 0.28, D= 0.26 L=2.8, D=2.6 L1= 3.32, L2= 3.32, D1= 2.83, D2= 3.13 L1= 0.688, L2= 0.632, D1= 0.604, D2= 0.485 L1= 0.054, L2= 0.042, D1= 0.061, D2= 0.039 L1= 6390.88, L2= 6014.00, D1= 6750.00, D2= 5115.59
mut2 (487) L= 0.24, D= 0.29 L= 2.4, D=2.9 L1= 3.28, L2= 3.27, D1= 3.33, D2= 3.32 L1= 0.643, L2= 0.560, D1= 0.526, D2= 0.459 L1= 0.009, L2= 0.012, D1= 0.008, D2= 0.007 L1= 7960.03, L2= 6875.00, D1= 5296.55, D2= 4644.78
Team Strain OD600 (1:10) OD600 Elapsed time (minutes) Abs420 (list both values) OD550 (list both values) Units
Blue WT
KD
mut1 (486) Light1 = 2.4, Dark1 = 2.9

Light2 = 2.4, Dark2 = 2.9

Light1 = 0.24, Dark1 = 0.29

Light2 = 0.24, Dark2 = 0.29

17 (across all samples) Light1 = 0.590, Dark1 = 0.598

Light2 = 0.572, Dark2 = 0.615

Light1 = 0.007, Dark1 = 0.008

Light2 = 0.009, Dark2 = 0.009

Light1 = 1419, Dark1 = 1184

Light2 = 1363, Dark2 = 1215

mut2 (487) Light1 = 2.8, Dark1 = 2.6

Light2 = 2.8, Dark2 = 2.6

Light1 = 0.28, Dark1 = 0.26

Light2 = 0.28, Dark2 = 0.26

17 (across all samples) Light1 = 0.650, Dark1 = 0.680

Light2 = 0.641, Dark2 = 0.679

Light1 = 0.006, Dark1 = 0.007

Light2 = 0.006, Dark2 = 0.007

Light1 = 1343, Dark1 = 1510

Light2 = 1324, Dark2 = 1508

Team Strain OD600 (1:10) OD600 Elapsed time (minutes) Abs420 (list both values) OD550 (list both values) Units
Pink WT
KD
mut1-487 L1&2-0.28, D1&2-0.26 L1&2-2.8, D1&2-2.6 L1&2-18.33, D1&2-17.67 L1-0.609, L2-0.616, D1-0.632, D2-0.621 L1-0.007, L2-0.003, D1-0.006, D2-0.006 L1-1162.7, L2-1190.0, D1-1352.8, D2-1328.8
mut2-488 L1&2-0.22, D1&2-0.25 L1&2-2.2, D1&2-2.5 L1&2-25.17, D1&2-25.17 L1-0.605, L2-0.607, D1-0.743, D2-0.756 L1-0.01, L2-0.005, D1-0.006, D2-0.006 L1-1061.0, L2-1080.4, D1-1164.1, D2-1184.7
Team Strain OD600 (1:10) OD600 Elapsed time (minutes) Abs420 (list both values) OD550 (list both values) Units
Purple WT
KD
mut1 (486) L: 0.18; D: 0.22 L: 1.8; D: 2.2 35 (for all) L1: 0.819; L2: 0.818; D1: 0.831; D2: 0.886; L1: 0.003; L2: 0.002; D1: 0.001; D2: 0.004 L1:1292.67; L2: 1292.86; D1: 1076.95; D2: 1141.56
mut2 (488) L: 0.22; D: 0.205 L: 2.2; D: 2.05 35 (for all) L1: 0.779; L2: 0.78; D1: 1.08; D2: 1.004 L1: 0.002; L2: 0.006; D1: 0.004; D2: 0.004 L1: 1007.14; L2: 999.35; D1: 1495.47; D2: 1389.55

|TR Purple Group Western Blot; Lane 1: H6 Envz, Lane 2: Wild Type, Lane 3: T541T Lane 4: 488- T541T

W/F b-gal data

Be sure to add the mutant strain # when you fill in this chart!

Team Strain OD600 (1:10) OD600 Elapsed time (minutes) Abs420 (list both values) OD550 (list both values) Units
Green WT D-1:.269, 2:.269, 3:.269, L-1:.273,2:.273,3:.273 D-1:21.5, 2:21.5, 3:21.5, L-1:21.5,2:21.5,3:21.5 D-1:.665, 2:.658, 3:.666, L-1:.305,2:.299,3:.286 D-1:.011, 2:.005, 3:.006, L-1:.013,2:.01,3:.009 D-1:1116.538, 2:1122.59, 3:1133.397, L-1:480.8757,2:479.5979,3:460.431
KD D-1:.226,2:.226,3:.226, L-1:.254,2:.254,3:.254 D-1:21.5,2:21.5,3:21.5, L-1:21.5,2:21.5,3:21.5 D-1:.036,2:.035,3:.037, L-1:.04,2:.04,3:.043 D-1:.005,2:.004,3:.006, L-1:.005,2:.005,3:.007 D-1:56.0815,2:57.62503,3:54.53797, L-1:56.56621,2:57.22395,3:56.30837
mut1 (487, T541Y) D-1:.23, 2:.23, 3:.23, L-1:.261,2:.261,3:.261 D-1:21.5, 2:21.5, 3:21.5, L-1:21.5,2:21.5,3:21.5 D-1:.717, 2:.771, 3:.788, L-1:.726,2:.772,3:.783 D-1:.01, 2:.01, 3:..011, L-1:..012,2:.012,3:.011 D-1:1414.56, 2:1523.761, 3:1554.601, L-1:.726,2:.772,3:.783
mut2 (488, T541P) D-1:.226, 2:.226, 3:.226, L-1:.252,2:.252,3:.252 D-1:21.5, 2:21.5, 3:21.5, L-1:21.5,2:21.5,3:21.5 D-1:.645, 2:.66, 3:.673, L-1:.671,2:.66,3:.64 D-1:.008, 2:.01, 3:.01, L-1:.01,2:.008,3:.011 D-1:.1298.621, 2:1322.289, 3:1349.043, L-1:1206.165,2:1192.322,3:1145.718
Team Strain OD600 (1:10) OD600 Elapsed time (minutes) Abs420 (list both values) OD550 (list both values) Units
Blue WT NB466 0.263, 0.255 (*light, dark*) 17.99, 18.00 0.221, 0.319 0.00433, 0.004 450.8, 1162.2 (stdev 11.7, 37.4)
KD NB468 0.249, 0.247 (*light, dark*) 17.99, 18.00 0.019, 0.026 0.0035, 0.00367 28.6, 53.8 (stdev 1.96, 2.72)
mut1 NB486 (T541R) 0.199, 0.199 (*light, dark*) 7.67, 7.67 0.266, 0.267 0.01, 0.007 1622.8, 1667.6 (stdev 128.6, 15.7)
mut2 NB488 (T541P) 0.257, 0.223 (*light, dark*) 8.67, 7.67 0.27, 0.253 0.005, 0.00467 1174.4, 1432.1 (stdev 20.5, 8.99)
Team Strain OD600 (1:10) OD600 Elapsed time (minutes) Abs420 (list both values) OD550 (list both values) Units
Pink WT
KD
mut1
mut2


WF Blue Group Western Blot (Mutant 1: no mutation, Mutant 2: T541S)

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