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Assessment
At all times wear a lab coat and suitable shoes with enclosed heel and toe and safety glasses
when required.
Always work to ensure your safety and the safety of those around you.
Microorganisms will be used in this class, avoid touching things to your mouth and wash hands
carefully after the class.
A risk assessment has been carried out for the practical classes and identified risks minimised. Please
observe the safety signs displayed and ask your demonstrator if you would like to know more, MSDS
( Material Safety Data Sheets) are available in the laboratory. Further information can be found at:
http://safety.unimelb.edu.au.
Time Management
Begin practical with the DNA extraction.
During Steps 1 and 3 incubations (each 15 minutes) start the analysis of the Arabidopsis plates.
DNA extraction should be completed at the end of one hour and shown to your demonstrator for
assessment.
Complete the observations of the Auxotroph.
P1- 2
BIOL10005: 2016
BIOL10005: 2016
P1 3
Equipment
A plastic tube containing E. coli suspension
Pipettes
Demonstrator initials
P1- 4
BIOL10005: 2016
Question 1:
Suggest why the solution may become viscous after the cell wall and cell
membrane are ruptured?
Question 2:
Question 3:
In a laboratory setting, the DNA you have extracted from E. coli would be purified and used in
other procedures such as gel electrophoresis (see practical 3)
BIOL10005: 2016
P1 5
Equipment
One pair of A. thaliana demonstration plates per pair of students
Procedure
1.
Question 4:
P1 =
F1 =
Plants of the F1 generation were allowed to self-fertilise and set seed (Figure 4). The F2 seed was
germinated.
You have been supplied with a petri dish containing young F2 plants.
2.
Score the glabrous/non-glabrous phenotype of the F2 plants and record your data and the
data from your bench in Table 1.
P1- 6
BIOL10005: 2016
Number of glabrous
(smooth) plants
Total
Own data
Class data
3.
Using the convention for Arabidopsis GL1 = non glabrous = hairy (wild type) and gl = glabrous
= smooth, propose a genetic hypothesis to account for these observations by filling in the
gaps below.
non glabrous
Parental
Genotypes
glabrous
H
H
F1
Genotype
non glabrous
H
h
Meiosis
F2
Female
gametes
Male
gametes
H
H
H
H
H
h
BIOL10005: 2016
Question 6:
Question 7:
P1 7
1. There is/ are ______ gene locus/i being considered in this cross with ______ allele/s. The
dominant phenotype is _________________________. The F2 of this _____________ cross will
produce a phenotypic ratio of ____________________.
2. State the null hypothesis:
Rarely do observed results exactly match the predicted results. Therefore we perform a
statistical test to see if the difference between the observed and the expected results is
statistically significant or not.
3. Using the appropriate statistical test (2) compare the predictions of your genetic model with the
observed results. Use the class data.
(Apart from the information in the BioByte, you can also refer to the Appendix 5.)
Calculate
the Chi-square (2) value for this comparison by completing Table 2 (for your
Table
2: 2 Analysis
calculations in genetic problems correct to 2 decimal places).
Phenotype
TOTAL
non glabrous (hairs)
glabrous (non hairy)
Observed
(O)
Expected
(E)
(O-E)2
E
2
(O-E)2
E
=
Use the Chi-square table (below).
Number of degrees of freedom
Probability
4. Conclusion: Circle the appropriate alternatives:
The null hypothesis is rejected / not rejected. Therefore, we accept / reject, the underlying genetic
hypothesis that gave rise to the prediction. That is, the observed results are consistent / not
consistent with the genetic hypothesis.
P1- 8
BIOL10005: 2016
NON-SIGNIFICANT
0.95
0.90
0.80
0.70
0.50
SIGNIFICANT
0.30
0.20
0.10
0.05
0.01
0.001
0.004
0.02
0.06
0.125
0.46
1.07
1.64
2.71
3.84
6.44
10.83
0.10
0.21
0.45
0.71
1.39
2.41
3.22
4.60
5.99
9.21
13.82
0.35
0.58
1.012
1.42
2.37
3.66
4.64
6.25
7.82
11.34
16.27
0.71
1.06
1.65
2.20
3.36
4.88
5.99
7.78
9.49
13.28
18.47
1.14
1.61
2.34
3.00
4.35
6.06
7.29
9.24
11.07
15.09
20.52
BIOL10005: 2016
P1 9
X
end of meiosis 1
end of
meiosis
2
F1
P1- 10
BIOL10005: 2016
BIOL10005: 2016
P1 11
Describe what you see on these 5 plates (hold the plate up to the ceiling to see
whether there is growth or not).
Question 10:
P1- 12
Question 11:
BIOL10005: 2016
Collect data for the 4 strains and enter the results in Table 5. Indicate when
growth occurred around the disc with a + and for no growth
Strain A
Strain B
Strain C
Strain D
Arg
Leu
Meth
Tyr
Phe
Water
Question 12:
a. Why is one disc impregnated with just water?
b. Had there been growth of E. coli around the disc impregnated with water, how would this
influence the conclusions you could draw from your experiment?
Question 13: Describe in what amino acid pathway there was a mutation for each of the strains.
Strain A
Strain B
Strain C
Strain D
Question 14: What was unusual about strain D? How can you explain this result?