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

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)