Drosophila Genetics Lab

Design Aspect 1: Define Problem/Hypothesis and Variables Problem or Research Question: When Sepia eye colored Drosophila is crossed with wildtype drosophila, what will be the phenotype outcome of the F2 generation? Hypothesis: When sepia drosophila is crossed with wildtype drosophila then the F1 generation will consist of all wildtype drosophila. When the F1 generation is crossed with itself, the F2 generation will consist of a 3:1 ratio of wildtype drosophila vs. sepia. Hypothesis Explanation: Assuming that the wildtype is dominant and sepia is recessive, the F1 generation will be completely filled of wildtype phenotypes because every fly will be heterozygous for sepia and the wildtype trait but will only express the dominant wildtype phenotype. However, when you cross a heterozygous generation with itself you get 1/4 homozygous for the dominant genotype, 1/2 heterozygous for the dominant and recessive genotype, and 1/4 homozygous for the recessive genotype. What this really means is that 3/4 of the flies will display the dominant phenotype since it will be displayed whether it has one or two alleles for the wildtype trait. Variables: Dependent: Phenotypes Independent: The Crossing of the flies Aspect 2: Controlling Variables Experimental Control: None Experimental Constants: -Amount of time allowed for flies to mate -Temperature -Amount of Medium -Number of flies in each vial Aspect 3: Method for Data Collection Apparatus and material: -Flies -Sorting Brushes -Sorting Cards -Microscope -Petri Dish -Fly Nap -Vial with medium Procedure: 1.In the parental generation we labeled on vial F1 generation and allowed the parentals to have offspring in that vial. 2.When the F1 generation flies hatched and mated, we used fly nap to anesthetize the flies. 3. The flies were put onto petri dishes with sorting cards under, using the aid of sorting brushes 4.Each individual fly was observed under a microscope to see its eye color and sex 5.We recorded the number of male and female flies along with their phenotype. 6.We then took the 13 female flies and 15 male flies, and put them in a vial named "F1 cross" 7.We allowed them to mate until the F2 generation eggs and larvae were able to be seen.

8.Fly nap was used to then knock out the F1 flies and they were disposed of, so as to not mate with the F2 flies. 9.When the F2 flies emerged we recorded their phenotype along with their sex. Data Collection and Processing Aspect 1: Recording Raw Data Qualitative data: Quantitative data: Table 1. Recording of F1 fly phenotype and sex (Sepia x Wildtype) Date Phenotype # of Females with # of Males with Total phenotype phenotype (Females + Males) Wildtype red eyes 13 15 28

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Table 2. Recording of F2 fly phenotype and sex (F1 Wildtype x F1 Wildtype) Phenotype Total Male Wildtype 28 Female Wildtype 36 Male Sepia 10 Female Sepia 10 Female Albino 1 Total: 85 Aspect 2: Processing Raw Data: Sample Calculations F2 Phenotype Ratio for Sepia x Wildtype # of Wildtype : # of Sepia # of Wildtype/# of Sepia Wildtype-64:Sepia-20 64/20=3.2 Wildtype-3.2:Sepia-1 Ratio=3.2:1 *Omission of albino anomaly Chi Squared Test Degree's of freedom=(Number of different Alleles-1)3-1=2 Chi squared= total of (observed-expected)^2/expected Expected is 3:1 ratio 64 observed wildtype, 20 observed sepia, 1 observed albino (64-75)^2/75 +(20-25)^2/25 + (1-0)^2/0 <-The albino is an unexpected value so it becomes undefined. =1.61+1=2.61<9.2 Value passes test

Aspect 3: Presentation of Processed Data: Types of Error There really wasn't much room for human systematic error as most of the experiment laid with the random processes of gene recombination. However, this does raise a point concerning our one albino fly. Under no presuppositions, the albino fly goes against all hypothesized outcomes. We were testing two genes, coding for either the wild type red eye or the mutant allele, sepia. Through our data we could conclude that the two genes were autosomal with wildtype red eyes remaining dominant while the sepia trait was simply recessive. How then, could we get a single drosophila with albino eyes? One hypothesis is that the trait was simply just genetic mutation, causing a slight change in the gene that coded for the eye color trait. There is, however, another plausible answer. Both sepia and red eyes are genes coding for eye color in drosophila, but there could, in fact, be another gene coding for eye color in general presupposing the actual color of the eye. This mechanism, is called epistatis, the phenomenon where the effects of one gene are modified by one or several other genes. So, in reality, the genes coding for either sepia or red eyes are still there but are being modified by another gene which is codes for either eye color, or no eye color. So what we are seeing in the albino fly is just the effect of epistasis, which actually skews our results a bit since we did not account for anything like this to happen at all. This also has the effect of adding one more degree of freedom to our chi square test even though the albino fly makes up only 1/85 flies. Discussion, Evaluation and Conclusion Aspect 1: Concluding Overall the data supports my hypothesis that when the F1 generation is crossed with itself, the F2 generation will consist of a 3:1 ratio of wildtype drosophila vs. sepia. This 3:1 ratio is a clear example of heterozygous crossing causing the phenotype expression of dominant vs. recessive traits. This also has the effect of suggesting that the wildtype is indeed dominant while sepia eye color is recessive. However, this realization would have been seen earlier without the full data, as the F1 generation which was a cross between a sepia and wildtype fly, consisted of flies with the wildtype phenotype. It was clear then that sepia could not have been a dominant trait, however, by examining the pattern of inheritance we could determine whether the genes were sex linked or autosomal. What we observed was that the traits popped up in male and female flies at pretty much the same rate ruling out possibility of sex linkage. For example, if sepia eye color had been a recessive sex linked trait on the X chromosome you would expect to see a greater proportion of male flies with sepia eye color versus female flies. Furthermore, through our chi squared test, the value we obtained was well within the range supporting our hypothesis. The anomaly in the experiment was caused by the occurrence of an albino drosophila. This skewed our data since we were only accounting for one gene the entire time, but what we didn't realize was that due to epistatis there were actually two genes to account for in eye color. While coding for color is still more dominant, there is still a chance that a fly can have albino eye color if the gene coding for color is turned off. So the albino coloring adds another degree of freedom to our chi squared test but because we were merely expected a 3:1 ratio there is no expected value for the albino so it becomes 0. During the test you must subtract the expected value from the observed and put it over the expected value. This causes a 0 in the denominator making the chi value for the albino undefined. Aspect 2: Evaluating Procedure(s) Overall this lab was pretty easy to execute and had pretty accurate results, however the glaring issue lies with a lack of control with the experiment. Sure we had multiple groups testing different crosses to obtain different variations but there was no standard to actually compare the crosses too. We did, however, have the chi squared test but that was based more on rejecting or accepting our hypothesis rather than comparing our variable experiment to a controlled experiment. Aspect 3: Improving the Investigation The addition of crossing another set of flies as a control group would be a welcomed addition to the experiment. I propose that along with crossing our sepia flies with the wildtype, that we also cross a separate wildtype x wildtype both homozygous in order to assure that we get wildtype flies in each generation. This would allow us a separate control group for us to actually compare our real crosses.