Talk:20.109(S12):Assess protein function (Day7)

Raw Data

Please post your raw calcium titration data below.

Day 7 data file, T/R	Day 7 data file, W/F
Media:S12_M2D7_TR-Red.txt	Media:S12_M2D7_WF-Red.txt
Media:S12_M2D7_TR-Orange.txt	Media:S12_M2D7_WF-Orange.txt
Media:S12_M2D7_TR-Yellow.txt	Media:S12_M2D7_WF-Yellow.txt
Media:S12_M2D7_TR-Green.txt	Media:S12_M2D7_WF-Green.txt
Media:S12_M2D7_TR-Blue.txt	Media:S12_M2D7_WF-Blue.txt
Media:S12_M2D7_TR-Pink.txt	Media:S12_M2D7_WF-Pink.txt
Media:S12_M2D7_TR-Purple.txt	Media:S12_M2D7_WF-Purple.txt

Analysis, T/R

Please be mindful of significant figures when you fill in this table. Do not state far more precise numbers than you can believe.

Group Colour	E67K, T79P, or M124S?	X#Z Mutant	Most believable KD; n for WT	Most believable KD; n for (+)	Most believable KD; n for X#Z mutant	Comments
Orange	T79P	D24H	0.104, 1.26	0.166, 1.30	0.7892, 2.73	I utilized model 2.
Purple	T79P	F65D	0.0848, 1.313	0.1329, 1.3673	0.1923, 0.6862	Utilized model 2
Blue	T79P	D57N	0.0993, 1.1927	.1036, 1.4481	.0613, 1.2909	Utilized model 2
Yellow	E67K	D20R	0.0907, 1.2464	0.0300, 0.8097	0.6699, 0.4722	Utilize model 2
Green	E67K	E67R	0.0635, 1.945	0.0202, 0.9558	0.0279, 1.1453	Utilized model 2
Red	E67K	E67R	0.1429, 1.3167	0.0384, 0.9833	0.0600, 1.0852	Model 2
Pink	T79P	F65D	0.0877, 1.2104	0.1333, 1.3360	0.4273, 1.2646	Model 2

Analysis, W/F

Please be mindful of significant figures when you fill in this table. Do not state far more precise numbers than you can believe.

Group Colour	E67K, T79P, or M124S?	X#Z Mutant	Most believable KD; n for WT	Most believable KD; n for (+)	Most believable KD; n for X#Z mutant	Comments
Orange	T79P	V55F	KD 0.1025; n 1.8227 for WT	KD 0.1491; n 1.3982 for T79P	KD 0.1598; n 1.6114 for V55F	The WT and T79P were switched during the experiment. For the above results, the labels were switched back to the original
Red	T79P	T79F	KD 0.1075; n 1.3550 for WT	KD 0.1372; n 1.1433 for T79P	KD 0.1053; n 1.3550 for T79F	As the above group, the labels were switched back to the original between T79P and WT. Also, our yield for WT and T79P was low (may be error prone)
Pink	E67K	D20K	KD 0.0986; n 1.2648 for WT	KD 0.0202; n 1.0027 for E67K	KD 0.7043; n 0.3610 for D20K	We used model 2 for our analysis. **Our data was updated in our revision process**
Purple	M124S	G23W/G25W	KD 0.1643; n 1.3591 for WT	KD 0.4753; n 2.9004 for M124S	KD 0.1470; n 1.0503 for G23W/G25W	Our yields for WT and G23W/G25W were low. The fluorescence values for G23W/G25W were very low (only between 18 and 24) and may not be significant. For G23W/G25W (contrary to WT and M124S), as [Ca2+] increased, fluorescence increased.
Blue	E67K	T26Y	KD 0.1047; n 1.2123	KD 0.0193; n 0.8686	KD 0.3547; n -0.6402	We used KD and hill coefficient values from model 2. Our T26Y had minimal variance between our highest (58 A) and lowest (47 A) absorbance values, contributing to some noise -- was also had a reversed transition curve.
Green	T79P	F65D	Method 1: KD 0.1151, Method 2:0.0872;Method 1: n 1.2658, Method 2: n = 1.1791	Method 1: KD 0.0969, Method 2:0.1018;Method 1: n 1.4356, Method 2: n = 1.6077	Method 1: KD 0.0599, Method 2:0.1523;Method 1: n 1.2302, Method 2: n = 1.4928	Results for both methods are as given.
Yellow	E67K	D20K	KD 0.2155; n 1.6616 for WT	KD 0.0556; n 1.0883 for E67K	KD 0.0910; n 0.2797 for D20K	We used the values from model 2. The Kd value and Hill coefficient for D20K should be ignored because from the raw data we can see that the fluorescence was basically flat, but high enough to know it's not background.