IBO 2013, SWITZERLAND Practical Exam “Molecular Cell Biology”
Student Code:
th
24 International Biology Olympiad
14th-21st July, 2013
Bern, Switzerland
Practical Exam 1
Molecular Cell Biology
Total points: 100
Duration: 90 minutes
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IBO 2013, SWITZERLAND Practical Exam “Molecular Cell Biology”
Dear participants,
This test consists of two tasks:
Introduction [2 points] 4
Task 1: Presence of the β-glucuronidase [12 points] 6
Part 1.1: Determine the presence of the β-glucuronidase [12 points] 6
Task 2: Presence of the procyclin protein [86 points] 7
Part 2.1: How to use the counting chamber [1 point] 7
Part 2.2: Washing the magnetic beads 8
Part 2.3: Density of trypanosomes not binding to magnetic beads [37.5 points] 8
Part 2.4: Total trypanosome density [25.5 points] 9
Part 2.5: Success of the deletion of the procyclin gene [9 points] 9
Part 2.6: Verification of binding to beads [9 points] 12
Part 2.7: Interpretation of your results [4 points] 13
Please write your student code into the box on the title page.
فقط جوابهاي داخل مربع. جوابها را فقط داخل مربعهاي خاکستري بنويسيد.پاسخنامه اي وجود ندارد
. حتما از اعداد انگليسي و خوانا استفاده کنيد. تصحيح با ماشين عالمت خوان انجام ميشود.تصحيح ميشود
7 و 1 به دليل اينکه تصحيح اعداد.جواب خود را به صورت تيک ( √ ) يا با اعداد انگليسي بنويسيد
. بنويسيد، در مربعهاي زير اين اعداد را به شکلي که معموالً يادداشت ميکنيد،بدون اشکال انجام شود
There is no separate answer sheet. Please fill in your answers into the specific answers boxes indicated
with a gray background. Only answers given inside these boxes will be evaluated.
The answers have to be given either with a tick (√) or with Arabic numbers. The numbers "1" and "7" can
look very similar in handwriting. To make sure that those two numbers can be well distinguished by the
IBO staff, please write them as you normally would into the following box.
1= 7=
Stop answering and put down your pen IMMEDIATELY when the bell rings at the end of the exam. Put
the entire protocol with all the answers back into the exam envelope.
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IBO 2013, SWITZERLAND Practical Exam “Molecular Cell Biology”
Material and equipment
Equipment
Water bath at 37°C (used in common)
1 Micropipette P1000
1 Micropipette P200
1 Micropipette P20
1 Box of pipette tips for P1000
1 Box of pipette tips for P200 and P20
1 Eppendorf holder
1 Tube holder
1 Solid waste container
1 Liquid waste tube [LW]
1 Polystyrene (styrofoam) box filled with ice
1 Timer
1 Marker
1 Microscope
3 Cell counting chambers
3 Microscope slides
3 Cover slips
21 Eppendorf tubes
1 Magnetic Eppendorf holder
Blank paper
1 Flag to call an assistant
1 Yellow sheet labeled with your student code
Chemicals
1 Eppendorf tube with magnetic beads [MB]
1 Tube with phosphate buffer [PBSB]
1 Eppendorf tube with Fixation Buffer [FB]
1 Eppendorf tube with Substrate Buffer [SB]
1 Eppendorf tube with Substrate (X-gluc) [S]
Trypanosome suspensions
1 Eppendorf tube with suspension of Strain 1 [T1]
1 Eppendorf tube with suspension of Strain 2 [T2]
1 Eppendorf tube with suspension of Strain 3 [T3]
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IBO 2013, SWITZERLAND Practical Exam “Molecular Cell Biology”
Introduction [2 points]
Trypanosoma brucei
Procyclin
GARP T. congolense
T. brucei brucei
GARP β-glucuronidase
β-glucuronidase
X-gluc T. brucei
X-gluc
111 GARP
A (magnetic beads)
Fc A
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IBO 2013, SWITZERLAND Practical Exam “Molecular Cell Biology”
. √
[2 points]
true false
If the suspension of a strain incubated with X-gluc turns blue, the
β-glucuronidase gene was successfully introduced.
Q1
X-gluc
Inferring the successful introduction of the GARP gene based on
the presence of the reporter gene (β-glucuronidase) may result in
false positives or negatives if only one gene of the construct was
successfully inserted.
GARP
The location in the genome where the construct is introduced
affects the level of gene expression of the introduced genes.
A similar approach with magnetic beads and specific antibodies
can be used to separate cells, with successful deletion of a gene
coding for an intracellular protein, from cells where the deletion
did not work.
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IBO 2013, SWITZERLAND Practical Exam “Molecular Cell Biology”
Task 1: Presence of β-glucuronidase [12 points]
Part 1.1: Determine the presence of β-glucuronidase [12 points]
T3 T2 T1 3
01 1
(SB) 111 0
(S) 11 3
37
yellow sheet
(√)
[12 points]
Strain T1 Strain T2 Strain T3
Q2
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IBO 2013, SWITZERLAND Practical Exam “Molecular Cell Biology”
Task 2: Presence of the procyclin protein [86 points]
Part 2.1: How to use the counting chamber [1 point]
0.2 0.3 counting chambers
0
11 1
0 0
2 3
40x 10x
1.1
[1 point] 0 1
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IBO 2013, SWITZERLAND Practical Exam “Molecular Cell Biology”
Volume of 1 counting square (mm³)
Q3
Volume of 1 counting square (ml)
Part 2.2: Washing the magnetic beads
0 MB
(PBSB) 1 1
0
(LW) 3
. 0 3
PBSB 33
Part 2.3: Density of trypanosomes not binding to magnetic beads [37.5 points]
T1 , T2 ,T3
191 1
11 0
3 3 31 3
0
2
3
3 (PBSB) 31 6
(PBSB) 11 1 111 7
FB 2 36 8
0.1 9
3 11
0.3
[37.5 points]
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IBO 2013, SWITZERLAND Practical Exam “Molecular Cell Biology”
Strain T1 Strain T2 Strain T3
Square 1
Q4
Square 2
Square 3
Mean counts per
square
Part 2.4: Total trypanosome density [25.5 points]
T1;T2, T3 3
PBSB 11 1 111 1
FB 2 36 0
0.1 3
2
[25.5 points] 0.3
̅ SD
√ ∑( ̅)
Strain T1 Strain T2 Strain T3
Square 1
Q5
Square 2
Square 3
Mean counts per square
Standard deviation of counts
per square
Part 2.5: Success of the deletion of the procyclin gene [9 points]
SEmean
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IBO 2013, SWITZERLAND Practical Exam “Molecular Cell Biology”
SEmean
√
0.2 SEmean
SEmean
Q6
The SEmean tells you the accuracy with which you estimated the mean number of trypanosomes per
square. Use this estimate to decide the correct number of significant digits (all digits including the first
for which you are uncertain) by comparing the estimated mean plus the SEmean with the estimated mean
minus SEmean. For instance, if the mean is 1234.567 and the SEmean 98.765, you will have to compare
1234.567 + 98.765 = 1333.332 with 1234.567 - 98.765 = 1135.802. In this case, there are two significant
digits and the mean should be reported as 1.2x103. Indicate the number of significant digits you should
use with your data. [1.5 points]
accuracy SEmean
SEmean SEmean
1234.567 + 98.765 = 1333.332 SEmean 98.765 1032.367
1.2x103 0 1234.567 - 98.765 = 1135.802
Number of significant digits
Q7
. [0.6 points]
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IBO 2013, SWITZERLAND Practical Exam “Molecular Cell Biology”
Strain T1 Strain T2 Strain T3
Mean counts per square of
trypanosomes not binding to magnetic
beads (from Part 2.3)
Q8
Mean counts per square of total
trypanosomes (from Part 2.4)
Now use these values to estimate the density of trypanosomes in the dilutions used for counting, and
report your values in the table below with the same number of significant digits. [3.7 points]
Strain T1 Strain T2 Strain T3
Trypanosomes not binding to
Q9 magnetic beads /ml in dilution used
for counting (from Part 2.3)
Total trypanosomes / ml in dilution
used for counting (from Part 2.4)
[1.1 points]
Strain T1 Strain T2 Strain T3
Trypanosomes not binding to
magnetic beads /ml in original
Q10
suspension (from Part 2.3)
Total Trypanosomes / ml in original
suspension (from Part 2.4)
In order to assess the success rate of the gene deletion experiment, calculate the percentage of
trypanosomes that did not bind to magnetic beads for each of the three strains. Use the estimates for
the densities in the original suspension for your calculations and indicate your results in the table below
(precision: only full percentages). [1.5 points]
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IBO 2013, SWITZERLAND Practical Exam “Molecular Cell Biology”
Strain T1 Strain T2 Strain T3
Percentage of trypanosomes not
Q11 binding to beads
Part 2.6: Verification of binding to beads [9 points]
6.2
trypanosomes
trypanosomes
0.3 11 1
0
[9 points] (√)
Strain T1 Strain T2 Strain T3
Practically no bound trypanosomes in the sample
Q12
<50% of the trypanosomes present in the sample are
bound
31
>50% of the trypanosomes present in the sample are
bound
31
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IBO 2013, SWITZERLAND Practical Exam “Molecular Cell Biology”
Part 2.7: Interpretation of your results [4 points]
(√)
[3 points]
Strain T1 Strain T2 Strain T3
A reduction, at least in part due to binding to
beads
Q13
No or only a purely stochastic change
(√)
Strain T1 Strain T2 Strain T3
Highest deletion efficiency
Q14
End of the Practical Exam
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