Biochemical Techniques- Flylab

GroupC01-Gr01
C01-Gr02
C01-Gr03
C01-Gr04
C01-Gr05
C01-Gr06
C01-Gr07
C01-Gr08
C01-Gr09
C01-Gr10
C01-Gr11
C01-Gr12
C02-Gr01
C02-Gr02
C02-Gr03
C02-Gr04
C02-Gr05
C02-Gr06
C02-Gr07
C02-Gr08
C02-Gr09
C02-Gr10
C02-Gr11
C02-Gr12
Strain
3976
3976
3976
3976
4062
4062
4062
4062
1118
1118
1118
1118
4062
4062
4062
4062
3976
3976
3976
3976
1118
1118
1118
1118
EMBA
Female/Male Total # of live flies
ST50
Male
5
13.25
Female
6
10
Male
6
15.5
Female
6
16
Female
9
14.5
Female
10
12.5
Male
7
13.5
Male
6
10
Male
8
20
Female
8
20
Male
6
20
Female
9
17.5
Male
6
17
Female
5
16.5
Female
6
24
Male
10
12.6
Male
10
14.5
Female
10
13
Male
8
14.5
Female
6
17
Male
10
22
Female
10
18
Male
6
17
Female
10
16
ST100
17
13
19
22
20
16
20
12
28
28
30
21
29
28
32
27
19
17
20
22
35
23
28
20
Extraction
# of flies
Volume (ul)
9
165
10
182
8
219
6
118
8
127
10
140
10
176
9
60
8
137
8
90
10
154
9
176
10
236
9
310
9
243
10
472
10
128
10
310
10
234
6
80
9
180
10
165
10
87
10
180
Activity assay
Dilution deltaA/min
0
0
0
0
0
0
0
0
5
0.03
0
0.036
0
0.037
0
0.019
0
0.034
0
0.053
0
0.041
0
0.028
3
0.053
0
0.012
0
0.017
0
0.058
0
0
0
0
0
0
0
0
0
0.031
0
0.019
0
0.021
0
0.029
Bradford assay
y=0.054x +0.1287
(DeltaA/min)/(6220M-cm- x 1cm)
Dilution
Absorbance protein concentraion mg/ml NAD+ molar concentration mol/L
0
0.556
7.912962963
0
0
0.368
4.431481481
0
0
0.387
4.783333333
0
0
0.332
3.764814815
0
0
0.531
7.45
4.82315E-06
2
0.416
5.32037037
5.78778E-06
0
0.377
4.598148148
5.94855E-06
0
0.471
6.338888889
3.05466E-06
0
0.611
8.931481481
5.46624E-06
0
0.491
6.709259259
8.5209E-06
0
0.401
5.042592593
6.59164E-06
0
0.328
3.690740741
4.50161E-06
0
0.401
5.042592593
8.5209E-06
0
0.352
4.135185185
1.92926E-06
0
0.547
7.746296296
2.73312E-06
0
0.374
4.542592593
9.32476E-06
0
0.258
2.394444444
0
0
0.426
5.505555556
0
0
0.314
3.431481481
0
2
0.219
1.672222222
0
0
0.423
5.45
4.98392E-06
0
0.475
6.412962963
3.05466E-06
0
0.279
2.783333333
3.37621E-06
0
0.363
4.338888889
4.66238E-06
molar con x 0.001 L x 10^6
NADH (umol) U
U/0.005ml
Relative activity x volume U/mg
Relative Activity U/mol Total Unit
Specific Assay
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0.004823151
0.964630225
122.5080386
16.44403202
0.005787781
1.15755627
162.0578778
30.45988654
0.005948553
1.189710611
209.3890675
45.53769491
0.003054662
0.610932476
36.65594855
5.782708799
0.005466238
1.093247588
149.7749196
16.76932544
0.0085209
1.704180064
153.3762058
22.86037845
0.00659164
1.318327974
203.022508
40.26153299
0.004501608
0.900321543
158.4565916
42.93354716
0.0085209
1.704180064
402.1864952
79.75788006
0.00192926
0.38585209
119.6141479
28.9259471
0.002733119
0.546623794
132.829582
17.14749564
0.009324759
1.864951768
880.2572347
193.7786004
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0.004983923
0.996784566
179.4212219
32.92132511
0.003054662
0.610932476
100.8038585
15.71876512
0.003376206
0.675241158
58.74598071
21.10634037
0.004662379
0.932475884
167.8456592
38.68401876
ST50 data from C01 & C02
sex
strain
male
female
13.25
10
15.5
16
14.5
13
14.5
17
3976
13.5
14.5
10
12.5
17
16.5
12.6
24
4062
20
20
20
17.5
22
18
17
16
1118
Anova: Two-Factor With Replication
SUMMARY male
female
Total
3976
Count
Sum
Average
Variance
Stdev
4
4
8
57.75
56
113.75
14.4375
14 14.21875
0.848958
10 4.704241
0.921389 3.162278 2.168926
4062
Count
Sum
Average
Variance
Stdev
4
4
8
53.1
67.5
120.6
13.275
16.875
15.075
8.369167 25.22917 18.10214
2.892951 5.022864 4.254661
1118
Count
Sum
Average
Variance
Stdev
4
4
8
79
71.5
150.5
19.75
17.875 18.8125
4.25 2.729167 3.995536
2.061553 1.652019 1.998884
Total
Count
Sum
Average
Variance
ANOVA
12
12
189.85
195
15.82083
16.25
12.33975 13.29545
Source of Variation SS
Sample
95.47896
Columns
1.105104
Interaction 32.22896
Within
154.2794
Total
283.0924
df
MS
F
P-value
F crit
2 47.73948 5.569835 0.013095 3.554557
1 1.105104 0.128934 0.723717 4.413873
2 16.11448
1.8801 0.181337 3.554557
18 8.571076
23
F is variance analysis
P-value is the most important
For P-value anything above 0.01 is
not accepted for sample.
Interaction mean any relationship
between ?
ST100 data from C01 & C02
sex
strain
male
female
17
13
19
22
19
17
20
22
3976
20
20
12
16
29
28
27
32
4062
28
28
30
21
35
23
28
20
1118
Anova: Two-Factor With Replication
SUMMARY male
female
Total
3976
Count
Sum
Average
Variance
Stdev
4
4
8
75
74
149
18.75
18.5
18.625
1.583333
19 8.839286
1.258306 4.358899 2.973094
4062
Count
Sum
Average
Variance
Stdev
4
4
8
88
96
184
22
24
23
59.33333 53.33333 49.42857
7.702813 7.302967 7.030546
1118
Count
Sum
Average
Variance
Stdev
4
4
8
121
92
213
30.25
23
26.625
10.91667 12.66667
25.125
3.304038 3.559026 5.012484
Total
Count
Sum
Average
Variance
ANOVA
12
12
284
262
23.66667 21.83333
45.15152 29.42424
Source of Variation SS
Sample
256.75
Columns
20.16667
Interaction 93.08333
Within
470.5
Total
840.5
df
MS
F
P-value
F crit
2 128.375 4.911265 0.019856 3.554557
1 20.16667 0.77152 0.391322 4.413873
2 46.54167 1.780553 0.196976 3.554557
18 26.13889
23
Source of Variati
strain
sex
female
0
0
0
0
0
0
0
0
45.53769491
16.44403202
5.782708799
30.45988654
79.75788006
28.9259471
193.7786004
17.14749564
16.76932544
22.86037845
40.26153299
42.93354716
32.92132511
15.71876512
21.10634037
38.68401876
male
3967
4062
1118
Anova: Two-Factor With Replication
SUMMARY
male
female
Total
3967
Count
Sum
Average
Variance
4
0
0
0
4
0
0
0
8
0
0
0
4062
Count
Sum
Average
Variance
Stdev
4
324.8568842
81.21422104
6545.195994
80.90238559
4
8
92.9773613 417.8342
23.24434033 52.22928
55.9201569 3789.195
7.477978129
4
111.0585239
27.76463098
115.9978773
10.77023107
4
8
120.1967095 231.2552
30.04917737 28.9069
165.8690053 122.2913
12.87901414
12
435.9154081
36.32628401
3055.899312
12
213.1740708
17.7645059
241.0404655
1118
Count
Sum
Average
Variance
Stdev
Total
Count
Sum
Average
Variance
ANOVA
Source of Variation
Sample
Columns
Interaction
Within
Total
SS
10953.17364
2067.237639
4664.214805
20648.9491
38333.57519
df
MS
F
P-value
F crit
2 5476.587 4.774023 0.021709 3.554557
1 2067.238 1.802042 0.196152 4.413873
2 2332.107 2.032933 0.159942 3.554557
18 1147.164
23
1118 (Female)
1118 (Male)
3976 (Female)
3976 (Male)
4062 (Female)
4062 (Male)
ST50 (avg.)
ST50 (stdev.) ST100 (avg.) ST100 (stdev.)
17.875
1.652019
23
3.559026
19.75
2.061553
30.25
3.304038
14
3.162278
18.5
4.358899
14.4375
0.921389
18.75
1.258306
16.875
5.022864
24
7.302967
13.275
2.892951
22
7.702813
Female
Strain 1118
Strain 3976
Strain 4062
Male
17.875
14
16.875
19.75
14.4375
13.275
MST decrease with increased ethanol concentration
25
MST (min)
20
15
10
5
0
Strain 1118
Strain 3976
Strain 4062
% ETOH
Female
Female
Strain 1118
Strain 3976
Strain 4062
Male
Male
23
18.5
24
30.25
18.75
22
MST dectease with increased ethanol concentration
40
35
MTS (min)
30
25
20
MTS (min)
20
15
10
5
0
Strain 1118
Strain 3976
Strain 4062
% ETOH
Female
Strain 1118
Strain 3976
Strain 4062
Male
Female
Male
30.04917737 27.76463098
0
0
23.24434033 81.21422104
Specific Activity
120
100
80
60
40
20
0
-20
Strain 1118
Strain 3976
-40
Female
Male
Strain 4062
Specific activity (U/mg) (avg.) Specific activity (U/mg) (stdev.)
30.04917737
12.87901414
27.76463098
10.77023107
0
0
0
0
23.24434033
7.477978129
81.21422104
80.90238559
Mahbubeh Arezomandi
BIBC 103
Fly Lab Report
You will present the data from your fly EMBA and ADH activity assays, and briefly
explain your findings and observations. The components that you will need to include in
your report is:
1. Data table (5 points): Refer to the “Template” tab of the “Fly lab data” sheet. Using the
class data fill out this table and include in your report. This data can be used to generate
the graphs below.
Strain
1118 (Female)
1118 (Male)
3976 (Female)
3976 (Male)
4062 (Female)
4062 (Male)
ST50 (avg.)
ST50 (stdev.) ST100 (avg.) ST100 (stdev.) Specific activity (U/mg) (avg.) Specific activity (U/mg) (stdev.)
17.875
1.652019
23
3.559026
30.04917737
12.87901414
19.75
2.061553
30.25
3.304038
27.76463098
10.77023107
14
3.162278
18.5
4.358899
0
0
14.4375
0.921389
18.75
1.258306
0
0
16.875
5.022864
24
7.302967
23.24434033
7.477978129
13.275
2.892951
22
7.702813
81.21422104
80.90238559
2. ST50 graph (10 points): Plot a female-male paired bar graph for three different genotypes
(1118, 3976, 4062) including the error bars with standard deviations. Perform 2-way
ANOVA on your samples with alpha 0.05 (default). Interpret your results based on the Pvalues. Are there any differences between samples (between different gender or
genotype)?
The p-value for the interaction is not significant (0.18134), so there is no interaction between the
3 strains or genotypes.
However, the p-value for the sample is significantly significant (0.013095), so we can say that
there are differences between samples.
The p-value for the columns is not significant (0.72372)
Strain 1118
Strain 3976
Strain 4062
Female
Male
17.875
19.75
14
14.4375
16.875
13.275
MST decrease with increased ethanol concentration
25
MST (min)
20
15
10
5
0
Strain 1118
Strain 3976
Strain 4062
% ETOH
Female
Male
3. ST100 graph (10 points): Plot a female-male paired bar graph for three different
genotypes including the error bars with standard deviations. Perform 2-way ANOVA on
your samples with alpha 0.05 (default). Interpret your results based on the P-values. Are
there any differences between samples (between different gender or genotype)?
Female
Strain 1118
Strain 3976
Strain 4062
Male
23
18.5
24
30.25
18.75
22
MST dectease with increased ethanol concentration
40
35
MTS (min)
30
25
20
15
10
5
0
Strain 1118
Strain 3976
Strain 4062
% ETOH
Female
Male
4. Conclusions of EMBA (10 points): The three genotypes of flies represent, sensitive,
resistant, and normal response to ethanol. Considering the ST50 and ST100 graphs, can
you infer anything about the different genotypes? Which one is sensitive/resistant to
ethanol, and which one has a normal response?
5. Specific activity graph (10 points): Plot a female-male paired bar graph for three different
genotypes (1118, 3976, 4062) including the error bars with standard deviations. Perform
2-way ANOVA on your samples with alpha 0.05 (default). Interpret your results based on
the P-values. Are there any differences between samples (between different gender or
genotype)?
Strain 1118
Strain 3976
Strain 4062
Female
Male
30.04917737 27.76463098
0
0
23.24434033 81.21422104
Specific Activity
120
100
80
60
40
20
0
-20
Strain 1118
Strain 3976
Strain 4062
-40
Female
Male
6. Conclusions of activity assays (5 points): The three genotypes of flies represent,
sensitive, resistant, and normal response to ethanol. Considering the specific activity
graph, can you infer anything about the different genotypes? Which allele is better at
breaking down alcohol? Are the results congruent with the EMBA results?