Malley 1

Rita Malley

Ms. Ferrara

AP Biology 4704-1

20 November 2008

Alternative to Lab 7: DrosophiLab

Introduction
The purpose of this lab was to study the basic principles of genetics through
experimenting with different kinds of drosophila melanogaster. In these experiments, we
simulated the basic principles of inheritance based on Mendelian genetics by performing
different crosses between different variations of fruit flies. Flies are used for this
experiment because they are prolific breeders, have a short generation time, have easy
characteristics to follow, such as different eye color, and have four chromosome pairs: 3
autosomes and 1 pair of sex chromosomes. For this lab, we used the program
DrosophiLab to simulate the genetics of fruit flies. To form our hypotheses for these
experiments, we used the hypothetic expected ratios predicted for Mendel’s monohybrid
cross and a dihybrid cross. When genetic crosses are actually performed, these crosses
are subject to random changes and experimental errors that produce chance deviation in
the actual phenotypic ratios that one may observe. In order to accurately evaluate genetic
inheritance, it is essential that observed deviation in a phenotypic ratio be determined and
compared with predicted ratios. Chi-square analysis is used to compare and evaluate the
data. This statistical method considers the random deviation for an observed ratio, and
the sample size of the offspring and expresses the data as a single value. In experiment 1,
we hypothesized that if the parents are true breeding for eye color, then the monohybrid
cross between a wild-eyed female with a sepia-eyed male will generate an F2 generation
with a 3:1 phenotypic ratio. In the second experiment, we predict that the pattern of
inheritance for this trait is sex-linked. We hypothesized that In the third experiment, the
cross is dihybrid and should follow the rules of simple Mendelian genetics. We
hypothesize that if the parental generation is true-breeding for both traits and the F1
generation is heterozygous for both eye color and wing type, then there the phenotypic
ratio will be 9:3:3:1 of red normal to red vestigial to sepia normal to sepia vestigial in the
F2 offspring for a simple dominance/recessive inheritance pattern for a dihybrid cross.
We will use the chi-square analysis to determine if any variance in data is significant and
to see if our hypothesis is correct.

Materials
DrosophiLab
Paper/pencil

General Procedure
 Use DrosophiLab to simulate crosses
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o Edit the genetic makeup using “chromosome editor” to get the desired
phenotype and genotype
o Select parents
o Generate F1 generation
o Select a male and female from F1 and set as parents for the F2 generation
o Cross parents from F1 to create F2 generation
o Evaluate and record results
 Calculate expected & observed values for F2 generation
o Expected: Study genotype of parents and create hypothesis using
principles of genetics
 Calculate expected values for each class
o Observed: record values observed based on F2 generation
 Calculate chi-square to evaluate how observed ratios for a given cross compare
with predicted ratios
o Use degree of freedom to determine if a variance in data is significant

Experiment 1: Monohybrid Cross

If the parents are true breeding for eye color, then the monohybrid cross between a wild-
eyed female with a sepia-eyed male will generate an F2 generation with a 3:1 phenotypic
ratio.
 Set up parent generation as a monohybrid cross between a wild-eyed female (red
eyes) with a sepia-eyed male
 Make the number of offspring 40 and show your results as “icons” (drag & drop)
 Record your results
 After this cross, save the offspring F1 in the flyfiles and perform a second cross,
F1 x F1 to obtain an F2 generation. Change the number of offspring to 1000 and
show results as “table”. Record your results.

Questions
1) What are the phenotypes of the parental generation and what is the nature of the
mutation (eye color, wind shape)?
a) In the parental generation, the phenotypes are: one parent has wild (red) eyes and
the other parent has sepia eyes; the nature of mutation in experiment one is eye
color.
2) What do the F1 generation indicate about the pattern of inheritance of this
character? Could this mutation be sex-linked? Explain your answer.
a) The F1 generation produces offspring with 2 different phenotypes: 75% of the
offspring will be wild eyed, while 25% will be sepia-eyed. This mutation is not
sex linked because the gene for eye color is not located on the sex chromosomes,
they are found on one of the fly’s autosomes. In the F1 offspring, all offspring
had wild eyes; they were heterozygous for the trait.
3) What are the genotypes of the F1 and P generations? Do a Punnett square for
the P and F1 generations.
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a) The genotypes of the P generation is one parent (male) homozygous recessive (rr)
and one parent (female) homozygous dominant (RR). The genotypes for the F1
generation is both parents heterozygous (Rr).
P generation: RR x rr

F1 Generation: Rr x rr

4) What are the expected phenotypic and genotypic ratios for the F1 and F2
generation?
a) The expected phenotypic ratio for the F1 generation is 1:1 wild eyed; the expected
genotypic ratio is 1:1 heterozygous for the eye color gene. The expected
phenotypic ratio for the F2 generation is 3:1 wild eyed to sepia eyed; the expected
genotypic ratio for the F2 generation is 50% heterozygous for wild eyes, 25% for
homozygous dominant for wild eyes, and 25% homozygous recessive for sepia
eyes. These predictions are based on the hypothesis that this monohybrid cross
follows the pattern of simple Mendelian genetics; a cross between a homozygous
dominant parent and a homozygous recessive parent produces a 1:1 ratio of
offspring that are heterozygous for the trait, which is the case in the F1
generation. In the F2 generation, the expected results also follow the pattern of
simple Mendelian genetics: a cross between two heterozygous parents yields a 3:1
phenotypic ratio (dominant to recessive) and a 2:1:1 ratio (heterozygous to
homozygous dominant to homozygous recessive).
5) Calculate the chi-square value for what you observed in the F2 generation. Show
all work. Is the variance in your data probably due to random chance alone and
therefore your hypothesis about the genetics of a trait is supported by the data?
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Phenotype O E O-E (O-E)2 (O-E)2

E
Sepia-eyed 261 1000 x .25 = 261-250 = (11)2 = (121)2 =
250 11 121 250
0.484
Wild eyed 739 1000 x.75 = 739- 750 = (-11) = 121 = 0.1613
750 -11 121 750
Total 1000 0.484+ 0.1613
= 0.6453
In this cross, the 5% probability from the table is 3.84 for 1 degree of freedom. My chi-
square value is 0.6453 and therefore less than 3.84. I can conclude that the variance in
my experimental data is not significant; therefore my hypothesis about the genetics of the
eye color trait is supported by the data from this cross.

Experiment 2:
If the trait for eye color is a monohybrid cross and the parents are true breeding, then
there will be a 1:1:1:1 ratio

 Set up your parent generation as a cross between a wild type male and a white
eyed female. Make the number of offspring 40 and show results as “icon”. Record
your results
 Save the F1 offspring in the flyfiles and perform a second cross F1 x F1 to obtain
F2 generational data. Change the number of offspring to 40 and show results as
“icon”. Record your results.
Questions
1) What do the results indicate about the pattern of inheritance for this character?
a) The results from this experiment indicate that the pattern of inheritance for this
character is sex-linked.
2) What are the expected phenotypic and genotypic ratios for the F1 and F2
generations?
a) In the F1 generation, the expected phenotype is 1:1 and the expected genotype is
1:1. In the F2 generation, the phenotypic ratio is 1:1:1:1 and the genotypic ratio is
9:3:3:1.
3) Calculate the Chi-square value for what you observed in the F2 generation. Show all
work. Is the variance in your data probably due to random chance alone and therefore
your hypothesis about the genetics of a trait is supported by the data?
Phenotype O E O-E (O-E)2 (O-E)2
E
Red male 13 10 3 (3)2 = 9 9 = 0.9
10
2
White male 8 10 -2 (-2) = 4 4_ = 0.4
10
2
Red female 7 10 -3 (-3) = 9 9_ = 0.9
10
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White 12 10 2 (2)2 = 4 4_ =0.4

female 10
Total 40 0.9+0.4+ 0.9
+0.4 = 2.6
The Chi-sqaure value for 1 degree of freedom is 3.841; therefore this hypothesis is
accurate. The variance in the data is due to random chance, because our value is less than
the critical value.

Third Experiment: Dihybrid cross

If the parental generation is true-breeding for both traits and the F1 generation is
heterozygous for both eye color and wing type, then there the phenotypic ratio will be
9:3:3:1 of red normal to red vestigial to sepia normal to sepia vestigial in the F2
offspring for a simple dominance/recessive inheritance pattern for a dihybrid cross.

 Cross a red vestigial female with a white normal winged male. Make your
number of offspring 1000 and show results in a “table”. Work through an F2
generation. Record all of your data.

Questions
1) What were the phenotypes and genotypes of your P, F1 and F2 generations?
a) For the parent generation, the phenotypes were one red, vestigial female and one
parent a sepia, normal winged male. The genotypes are homozygous dominant
for red eyes and homozygous recessive for vestigial wings for the female parent;
for the male, homozygous recessive for sepia eyes and homozygous dominant for
normal winged.
b) For the F1 generation, the phenotypes of all the offspring were the same: red eyed
and normal winged. The genotype was also the same for all offspring:
heterozygous for both traits.
c) For the F2 generation, the pheotypes of the offspring were red-eyed vestigial, red-
eyed normal winged, sepia-eyed vestigial, and sepia-eyed normal winged. The
genotypes of the F2 generation were:
i) Homozygous dominant for both traits
ii) Homozygous dominant for eye color, heterozygous for wings
iii) Heterozygous for eyes, homozygous dominant for wings
iv) Heterozygous for both traits
v) Homozygous recessive for both traits
vi) Homozygous dominant for eye color, homozygous recessive for wings
vii) Heterozygous for eye color, homozygous recessive for wings
viii) Homozygous recessive for eye color, homozygous dominant for wings
ix) Homozygous recessive for eye color, heterozygous for wings
2) What were the expected phenotype ratios for your generations?
a) The expected phenotype ratios for the F2 generation was 9:3:3:1; we predicted
that 56% of the offspring would be red eyed and normal winged, 19% would be
red eyed and vestigial winged, 19% would be sepia eyed and normal winged, and
6% would be sepia eyed and vestigial wings.
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3) Calculate the chi-square value for what you observed in the F2 generation. Show
all work.

Phenotype O E O-E (O-E)2 (O-E)2

E
Red & 586 563 23 (23)2 = 529 529 = 0.9027
normal 586
Red & 179 187 -8 (-8)2 = 64 64_ = 0.3575
vestigial 187
Sepia & 169 187 -18 (-18)2 = 324 324_ 1.9172
normal 169
Sepia & 66 63 3 (3)2 = 9 9_ = 0.1364
vestigial 66
Total 1000 0.9027+
0.3575+1.9172
+ 0.1364 =
3.3138

Questions:
1) What is the chi-square test used for?
a) Chi square is a statistical tool that is used to compare genetic data obtained
through experimentation with hypothetical ratios for the suspected pattern of
inheritance. This test is used by genetics to evaluate data from experimental
crosses to determine if the assumed genetic explanation is supported by the data.
It helps the experimenter to decide if the difference between your observed results
and your expected results is probably due to random chance alone, or if there is
some other factor influencing the results.
2) Why is probability important in genetics?
a) In genetics, probability estimates the likelihood of gene distribution from one
generation to the next. Most eukaryotic organisms are diploid, meaning that each
cell contains two copies of every chromosome, and for each gene on a
chromosome, there are two alleles that controls a trait. In sexual reproduction,
these two copies of each chromosome separate, and are randomly sorted into the
gametes. When gametes from two different parents combine in fertilization, new
combinations of these alleles are made. Probability determines which alleles, and
therefore which combinations of characteristics is the result of random chance.
3) Briefly describe how the Chi-square analysis may be used in genetics.
a) In genetics, the chi-sqaure analysis can be used to support or nullify hypotheses
and predications about the pattern of inheritance for certain traits. Comparing
data to the critical values from Chi-square show if the variances in data are from
random chance or from an incorrect hypothesis.
4) Suppose you were to obtain a chi-square value of 7.82 or greater in your data
analysis (with 2 degrees of freedom). What would this indicate?
a) A chi-square value of 7.82 or greater would indicate that the variance in data is
significant; the difference between the expected values and observed values is not
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due to chance alone. Therefore, this value of chi-square does not support the
hypothesis made because the variance is too great to be insignificant.
5) Suppose you were to obtain a Chi-square value of 4.60 or lower in your data
analysis (with 2 degrees of freedom). What would this indicate?
a) Because the critical value for 2 degrees of freedom at 5% is 5.991, a chi-square
value of 4.60 or lower means that the variance in data is not significant. This
value supports the hypothesis made because the variance falls within the range for
5% at 2 degrees of freedom.
6) A heterozygous white-fruited squash plant is crossed with a yellow-fruited plant,
yielding 200 seeds. Of these, 110 produce white-fruited plants while only 90
produce yellow fruited plants. Are these results statistically significant? Explain
using chi-square analysis. What is there were 2000 seeds and 1100 produced
white-fruited plants & 900 yellow-fruited?

Phenotype O E O-E (O-E)2 (O-E)2

E
White 110 100 10 (10)2 = 100 100 = 1
fruited (Ff) 100
Yellow 90 100 - 10 (10)2 = 100 100_ = 1
fruited (ff) 100
Total 200 1+1=2
For the first cross that yields 200 seeds, the variance in data is insignificant because the
value is less than 3.841, which is the critical value of the Chi-square distribution for 1
degree of freedom. These results support the hypothesis made about the first cross; any
variance in data is due to random chance alone.

Phenotype O E O-E (O-E)2 (O-E)2

E
White 1100 1000 100 (100)2 = 10000 = 10
fruited (Ff) 10000 1000
Yellow 900 1000 - 100 (10)2 = 10000 10000_ = 10
fruited (ff) 1000
Total 2000 10 + 10 = 20

For the second cross that yields 2000 seeds, the variance in data is significant because our
value is 20, while the critical value for 1 degree of freedom is 3.841. These results do not
support the expected results predicted according to the hypothesis, so the hypothesis is
invalid.

Conclusion
In this lab, we studied the basic principles of genetics through the simulation of 3
different crosses of fruit flies, drosopila melanogaster. In the first experiment, we
performed a monohybrid cross between a wild-eyed female and a sepia-eyed male. We
found that the 100% of the offspring in the F1 generation were heterozygous for eye
color; therefore, all showed the dominant trait (wild eyes). In the F2 generation, we
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found that there was an approximate 3:1 phenotypic ratio (wild eyes to sepia eyes).
Using the chi-square analysis, we found that our data supports hypothesis because our
value (2.6) was less than the critical value for Chi-square at 1 degree of freedom (3.841).
In the second experiment, we crossed a wild type male with a white type female; we
found that the trait for eye color is sex-linked. Our hypothesis that –the phenotypic ratio
for this experiment is 1:1:1:1 is incorrect because when we cas supported by our data
after we performed the chi-square analysis; our value of 2.6 is less than the critical value
on the chi-square chart, which is 7.815 for 3 degrees of freedom. In the third
experiment, we performed a dihybrid cross between