AP Biology Lab Name: _____________________________

Fly Lab - Genetics of Organisms

Objectives

Before doing this lab you should understand:

Chi-square analysis of data & the life cycle of diploid organisms useful in genetics studies.

After doing this lab you should be able to:

Investigate the independent assortment of two genes and determine whether the two genes are
autosomal or sex-linked using a multigenerational experiment, and analyze the data from your
genetic crosses using chi-square analysis techniques.

Statistical Analysis Section

Example 1

Statistics can be used to determine if differences among groups are significant, or simply the
result of predictable error. The statistical test most frequently used to determine whether data
obtained experimentally provide a good fit or approximation to the expected or theoretical data is the
chi-square test. This test can be used to determine if deviations from the expected values are due to
chance alone, or to some other circumstance. For example, consider corn seedlings resulting from an
F1 cross between parents that are heterozygous for color.

A Punnett square of the F1 cross Gg x Gg would predict the expected proportion of green:albino
seedlings would be 3:1. Use this information to fill in the Expected (e) column and the (o-e) column in
Table 7.1.

Table 7.1
Phenotype Genotype Observed (o) Expected (e) (o-e)
Green GG or Gg 72
Albino gg 12
Total: 84

There is a small difference between the observed and expected results, but are these data close enough
that the difference can be explained by random chance or variation in the sample?

To determine if the observed data fall within the acceptable limits, a chi-square analysis is performed
to test the validity of a null hypothesis (that there is no statistically significant difference between the
observed and expected data). If the chi-square analysis indicates that the data vary too much from the
expected 3:1, an alternative hypothesis is accepted.

The formula for chi-square is 2 =  (o-e)2

e
where o = observed number of individuals
e = expected number of individuals
 = the sum of values (in this case the differences, squared, divided by the number of expected)

1. This statistical test will examine the null hypothesis, which predicts that the data from the
experimental cross above will be expected to fit the 3:1 ratio. Write your null hypothesis below.

2. Use the data from Table 7.1 to complete table 7.2.

Table 7.2
Phenotype Observed (o) Expected (e) (o-e) (o-e)2 (o-e)2
e
Green 72
Albino 12

2 = (o-e)2
e

Critical Values Table

Degrees of Freedom (df)
Probability (p) 1 2 3 4 5
0.05 3.84 5.99 7.82 9.49 11.1
0.01 6.64 9.21 11.3 13.2 15.1
0.001 10.8 13.8 16.3 18.5 20.5

How to use the Critical Values Table

1. Determine the degrees of freedom (df) for your experiment. It is the number of phenotypic classes
minus 1. Since there are two possible phenotypes, for this experiment df=1 (two samples -1). If this
experiment had gathered data for a dihybrid cross, there would be four possible phenotypes and
therefore 3 degrees of freedom.

2. Find the p value. Under the 1 df column, find the critical value in the probability (p) = 0.05 row; it
is 3.84. What does this mean? If the calculated chi-square value is greater than or equal to the
critical value from the table, then the null hypothesis is rejected. Since in our example 2 =
5.14 and 5.14 > 3.84, we reject our null hypothesis that there is no statistically significant
difference between the observed and expected data. In other words, chance alone cannot explain
the deviations we observed and there is, therefore, a reason to doubt our original hypothesis (or to
question our data collection accuracy.) The minimum probability for rejecting a null hypothesis is
generally 0.05, so this is the row to use in s chi-square table.

3. These results are said to be significant at a probability of p = 0.05. This means that only 5% of the
time would you expect to see similar data if the null hypothesis was correct; thus, you are 95%
sure that the data did not fit a 3:1 ratio.

4. Since these data do not fit the expected 3:1 ratio, you must consider reasons for this variation.
Additional experimentation would be necessary. Perhaps the sample size was too small, or errors
were made in data collection. In this example, perhaps, the albino seedlings are underrepresented
because they died before the counting was performed.

Example 2
In a study of incomplete dominance in tobacco seedlings, the counts in table 7.3 were made from a
cross between two heterozygous plants. Complete the table below.

Table 7.3
(o-e)2
Phenotype Observed (o) Expected (e) (o-e) (o-e)2
e
Green 22
Yellow Green 50
Albino 12

2 = (o-e)2
e

A Punnett square for this cross indicates an expected ratio of 1 green:2 yellow green:1 albino.
Calculate the expected for each and fill out the Table 7.3.
How many degrees of freedom are there? _______

What is the probability value? ________ What is the critical value? _______________

According to the critical value of 2, do you fail to reject or reject the null hypothesis (does the data fit
the expected 1:2:1 ratio?)

Practice Problem

An investigator observes that when pure-breeding, long-winged fruit flies are mated with pure-
breeding, short-winged flies, the F1 offspring have an intermediate wing length. When several
intermediate-winged flies are allowed to interbreed, the following results are obtained:

230 long-winged flies, 510 intermediate winged flies, 260 short winged flies

a. What is the genotype of the F 1 intermediate-wing-length flies?

b. Write a hypothesis describing the mode of inheritance of wing length in fruit flies (this is your null
hypothesis).

c. Complete Table 7.4

Table 7.4
(o-e)2
Phenotype Observed (o) Expected (e) (o-e) (o-e)2
e

2 = (o-e)2
e

d. How many degrees of freedom (df) are there? _____

e. Chi-square (2) = _____

f. What is the probability value? ________ What is the critical value? _______________

g. Do you fail to reject or reject the null hypothesis? Explain why.

Procedure: Monohybrid Cross

1. Go to the website: https://www.sciencecourseware.org/FlyLabJS/

2. Begin your investigation into fruit fly genetics by choosing a male wild type for wing size and a
female with vestigial wing size.
3. Mate the flies and record the Data in Table 7.5.

Table 7.5-F1 Generation

Phenotype and Symbol Females Males
4. Select an F1 female and an F1 male to mate. Mate these flies and record the F2 generation data in
Table 7.6

Table 7.6-F2 Generation

Phenotype and Symbol Females Males

5. Write a hypothesis that describes the mode of inheritance of the trait you studied. This is your null
hypothesis.

6. Construct Punnett squares to predict the expected results of both parental and F 1 generational
crosses from your null hypothesis.

Parental Cross F1 Cross

7. Refer to the Punnett squares above. In the table below, record the expected ratios for the genotypes
and phenotypes of the F1 and F2 crosses in the experiment.

Expected Genotypic Ratio Expected Phenotypic Ratio

F1 generation
F2 generation

8. From the results, describe your cross.

Is the mutation sex-linked or autosomal? _________________

Is the mutation dominant or recessive? _________________

Is the cross monohybrid of dihybrid? _________________

9. Perform a chi-square test on your F2 results to see if the deviations are within limits expected by
chance.
 (o-e)2
Phenotype Observed (o) Expected (e) (o-e) (o-e)2
e

2 = (o-e)2
e
10. Calculate the chi-square value for these data.

How many degrees of freedom are there? _______________

Chi-square (2) = ____________

What is the probability value? ________ What is the critical value? _______________

11. Do you fail to reject or reject your null hypothesis? Explain.

Procedure: Monohybrid Cross

1. Return to the original list of fly choices.

2. Choose a male wild type and a female with a yellow body.

3. Mate the flies and record the Data in Table 7.7.

Table 7.7-F1 Generation

Phenotype and Symbol Females Males

4. Choose one female and one male from the F 1 Generation to mate. Mate them and record the F2
generation data in Table 7.8

Table 7.8-F2 Generation

Phenotype and Symbol Females Males
5. Write a null hypothesis that describes the mode of inheritance of the trait you studied.

6. Construct Punnett squares to predict the expected results of both parental and F 1 generational
crosses from your null hypothesis.
7.
Parental Cross F1 Cross

8. Refer to the Punnett squares above. In the table below, record the expected ratios for the genotypes
and phenotypes of the F1 and F2 crosses in the experiment.

Expected Genotypic Ratio Expected Phenotypic Ratio

F1 generation
F2 generation

9. From the results, describe your cross.

Is the mutation sex-linked or autosomal? _________________

Is the mutation dominant or recessive? _________________

10. Perform a chi-square test on your F2 results to see if the deviations are within limits expected by
chance.

Phenotype Observed (o) Expected (e) (o-e) (o-e)2 (o-e)2

e

2 = (o-e)2
e

11. Calculate the chi-square value for these data.

How many degrees of freedom are there?

Chi-square (2)

What is the probability value? ________ What is the critical value? _______________
12. Can you accept or reject your null hypothesis? Explain.

Procedure: Dihybrid Cross

1. Return to the original list of fly choices.

2. Select a wild type female and a male with sepia eyes and black body.
3. Mate the flies and record the Data in Table 7.9.

Table 7.9-F1 Generation

Phenotype and Symbol Females Males

4. Select a female and a male from the F1 Generation to mate. Mate these flies and record the F2
generation data in Table 7.10

Table 7.10-F2 Generation

Phenotype and Symbol Females Males

5. Write a null hypothesis that describes the mode of inheritance of the traits you studied.

6. Construct Punnett squares to predict the expected results of both parental and F 1 generational
crosses from your null hypothesis.

Parental Cross F1 Cross

7. Refer to the Punnett squares above. In the table below, record the expected ratios for the genotypes
and phenotypes of the F1 and F2 crosses in the experiment.

Expected Genotypic Ratio Expected Phenotypic Ratio

F1 generation
F2 generation

8. From the results, describe your cross.

9.
Are the mutations sex-linked or autosomal? _________________

Are the mutations dominant or recessive? _________________

Is the cross monohybrid of dihybrid? _________________

10. Perform a chi-square test on your F2 results to see if the deviations are within limits expected by
chance.
Phenotype Observed (o) Expected (e) (o-e) (o-e)2 (o-e)2
e

2 = (o-e)2
e

11. Calculate the chi-square value for these data.

How many degrees of freedom are there?

Chi-square (2)

What is the probability for this data?

12. Can you accept or reject your null hypothesis? Explain why?