*This is one of my old college lab reports!

Forest Thomer
Introduction

Mendel proposed the Law of Independent Assortment. This law is in relation to the known fact that mutant genes are produced in frequencies that result from random movement in meiotic chromosome pairs. According to this law the alleles of different genes assort independently. Yet this law only applies when the loci are on different pairs of chromosomes. A linkage gene would describe mutants located on the same homolog (chromosome pair) (Thompson 2006).

Thus the purpose of this experiment was to tangibly determine if the recessive mutant genes for apterous wings and Sepia eye color (black) are linked in the diptera species Drosophila melanogaster or commonly referred to as the fruit fly. Since the experiment aims to prove that the genes are linked the null hypothesis would imply there to be no significant difference between what is observed and what is expected, if and only if the genes are linked. If the genes are linked we can expect a 2, 1, 1, 0 ratio. This meaning 2/4 of the cultivated flies will not exhibit either recessive trait, ¼ will exhibit apterous wings and red eyes, ¼ will have normal wild type wings and sepia eyes, and none will have the presence of both recessive traits due to them being physically located on the same chromosome. If the genes are not linked we can expect a 9,3,3,1 ratio in which 9/16 of the cultivated flies will not exhibit either recessive trait, 3/16 will exhibit apterous wings and red eyes, 3/16 will have normal wild type wings and sepia eyes, and 1/16 will have the presence of both recessive traits due to crossing over, thus demonstrating them being located on different chromosomes.

Materials
For the experiment to be fulfilled three vials of various genotypic and phenotypic Drosophila melanogaster were provided by the instructor. One vial included nothing but Wild Type flies. Thus their eye color was expressed as red and they had the presence of wings. The second vile had sexually diverse members, whom were homozygous recessive for the presence of no wings (apterous) while having red wild type eyes. The third vile included sexually diverse members of the species whom were homozygous recessive for the presence of sepia (dark/black) eye color and exhibited the wild type (long) wing characteristic. The flies were to be considered as such since the “provider” was a specialized carrier of the specimens.
Separate autoclaved vials were prepared using a commercially prepared, instant type of medium. The medium was prepared by placing a “standard measure” of dry medium and about 15mL of water into autoclaved vials. The mixture was swirled to remove air bubbles and then allowed to dry for two minutes. A sterilized sponge stopper was used to prevent further contamination and as a barrier to prevent the specimens from escaping once introduction had occurred. CO2 gas was used to facilitate cultivation as were horse haired brushes. A microscope was provided in order to aid sexual identification and also make recessive trait “expression” more accurate. (I will not provide a diary of activities concerning fruit fly culturing because such a description is not necessary for this lab report).

Once desired virgin female flies were cultivated a parental cross involving thirteen (13) male flies and thirteen (13) female flies was made on 10/29/06. The parental cross involved thirteen males that exhibited sepia eye color and the wild type (present) wing. This cross also involved thirteen virgin female flies which exhibited wild type eye color, red eyes, and apterous (the lack of wings) wing type. It is crucial to cultivate virgin females in order to guarantee the results are accurate. This is the case because females store sperm from previous partners. In order to “ensure” virgin female flies are cultivated, cultivation must be done within 2-4 hours after the fly ecloses/emerges from their pupae. Other precautions include clearing adults from the vial once a “significant” and/or reasonable number of pupae appear (roughly 5-6 days after parental cross has been made). On 11/6/06 and 11/9/06 the F1 generation was counted. The F2 generation was introduced into a sterile vial on 11/6/06. The F2 generation was then counted on 11/13/06 and 11/20/06. The F2 generation was produced by taking ten (10) males that exhibited sepia eye color and the wild type (present) wing and ten (10) female flies which exhibited wild type eye color, red eyes, and apterous (the lack of wings) wing type. All fly crosses were done in a vial which was placed inside an incubator set at 27 degrees Celsius. A Drosophila melanogaster life cycle diagram is located at figure 1.
FIGURE 1

http://media.nasaexplores.com/lessons/04-006/images/life-cycle.gif

Results

The following diagrams document the observed flies from both the F1 and F2 generation cultivated on the mentioned dates.

F1 Generation

Phenotypes Male Female Total

11/6/06.………11/9/06 11/6/06.……11/9/06
Long wings, red eye color 9 28 6 42 85
Apterous wings, red eye color 0 0 0 0 0
Long wings, sepia eye color 0 0 0 0 0
Apterous wings, sepia eye color 0 0 0 0 0
Totals 37 48 85

F2 Generation

Phenotypes Male Female Total

11/13/06.……11/20/06 11/13/06.……11/20/06
Long wings, red eye color 2 63 34 96 195
Apterous wings, red eye color 2 14 0 0 0
Long wings, sepia eye color 0 0 0 0 0
Apterous wings, sepia eye color 0 0 34 96 0
Totals 81 130 211

A Chi-Square Test was preformed using the F2 Generation data. A Chi-Square Test is a commonly used basic statistical tool used to compare observed experimental results with the values that are theoretically to be expected (Thompson 2006). This test involved the use of a Chi-Square diagram handed out in class in order to determine the p-value. A p-value is the probability of rejecting the null hypothesis (that there is no difference between observed and expected), when the null hypothesis is true. In other words the p-value gives an approximate value for the error of falsely stating that there is a significant difference between your observed numbers and the expected numbers (Carter 2002). This test was conducted to determine the statistical probability/likely-hood of two different null hypotheses. The null hypothesis used that tested genes as linked was as follows…

-There is no significant difference between what we observed and what is expected if the genes are linked.

The null hypothesis that tested genes as not linked was as follows…

-There is no significant difference between what we observed and what is expected if the genes are not linked.

This statistical test requires taking the sum of (observed-expected) ^2/expected (formula used for conducting this statistical test) for each expected class. Observed is the total number of flies observed for a class, for example 195 flies with wild type (long) wings and red eyes were observed in that class. We then take the number of observed flies and subtract the number of expected flies. The number of flies expected varies depending on which null hypothesis we are testing. As mentioned before, if the genes under examination are linked then we can expect a 2, 1, 1, 0 ratio. This meaning 2/4 of the cultivated flies will not exhibit either recessive trait, ¼ will exhibit apterous wings and red eyes, ¼ will have normal wild type wings and sepia eyes, and none will have the presence of both recessive traits due to them being located on the same chromosome. If the genes are not linked we can expect a 9,3,3,1 ratio in which 9/16 of the cultivated flies will not exhibit either recessive trait, 3/16 will exhibit apterous wings and red eyes, 3/16 will have normal wild type wings and sepia eyes, and 1/16 will have the presence of both traits due to them being located on different chromosomes.

We take the mentioned rations and multiply them with the total number of flies observed. For example…

If the genes are linked and we are trying to determine the expected (e) we would take 2/4 for the first class (not exhibiting either recessive trait) and multiply it with 211 (the total number of flies cultivated). The 2/4 ratio is used simply due to the “assumption” the genes are linked. Other statistical tests which have been explained in class have proven such a pattern, hence the reason the ratios used are being used. The resulting number is a statistical expectation in reference to the number of flies to be cultivated that fit the class description.

The (e) value for each class will be determined in that fashion. Upon finding the difference between observed and expected, we than square the resulting number and divide it by the class expected (e) value.
Once this has been determined and the sums of each equation have been added up for all the involved classes, an X^2 (Chi-Square) value has been found. Using a Chi-Square sheet handed out in class, the probability of this particular X^2 value was to be determined. In order to accomplish this, the degrees of freedom had to be determined simply to use the chart. This value is found by subtracting one from the number of expected classes. Thus 2 was the degrees of freedom used to test the null hypothesis dealing with the genes being linked (three expected classes minus one) and 3 was the degrees of freedom used to test the null hypothesis dealing with the genes not being linked (four expected classes minus one).

Discussion

The tested null hypotheses were rejected. As explained in class the genes are not linked. The mutant gene for sepia eye color appears on chromosome number (III) 3 and the mutant gene for apterous wings appear on chromosome number (I) 1 (Raymond 1988). Thus from a “tangible/visual” perspective the genes are not linked/found on the same chromosome. This means that crossing over should have occurred. The expected phenotypic ration of 9/16, 3/16, 3/16, and 1/16 should have been observed. Yet the genetic recombination observed in the F2 generation never produced any organisms with the wild type long wing and the mutant recessive gene for sepia eye color. According to the statistical results both the X^2 values were less than .05 (the critical value). The X^2 (Chi-Square) value testing the linked gene hypothesis equaled 135.33 and 115.75 was the ending sum testing the not linked hypothesis. These numbers were used to determine p-values. According to the given chart, these numbers indicate reason to doubt the hypotheses because the observed data differed significantly from the expected. As such “implies” the difference cannot be due only to chance sampling variation. Had the statistic been greater than the critical value the null hypothesis would have not been rejected? Reasons for this outcome range from numerous potential errors. It is possible I did not count and or observe all the flies in the vials. Flies often “escape” despite best efforts to contain them. Thus I may have had flies presented with wild type long wings and the recessive mutant gene for sepia eye color. It is very likely they flew away. It is possible that I may have done my count too early. Some flies were still in their pupae stage and thus yet to eclose. If the count was done two days later maybe different ratios would have been observed supporting one of the null hypotheses. Other error could have been the results of poor cultivating on my part. One could assume that the cultivated “virgin” females may very well not been virgin. This issue is of course one of the harder ones to address simply because the current lab and staff hours, school schedules, work schedules, etc. do not permit myself or anyone else to monitor the flies twenty-four hours daily until the experiments conclusion. Counting the cultivated flies may have been interrupted by more urgent issues (such as a class, ride home, etc.) thus making one careless and not as observant as they would normally be. Flies could have “hid” themselves under shed exoskeleton, the media, etc. thus it is possible all the adult flies were not removed from the vials after each count. Thus offspring could have been reproducing with parent, this in turn not providing adequate phenotypic expression. Numerous events could have, and most likely did occur, that generated the data presented. But the data is the data. I can not change the events of the past hence my observations are all that I can work with. More experimentation is recommended to determine if the null hypotheses truly should be rejected.

Conclusion
According to the data obtained in this experiment the null hypothesis that, there is no significant difference between what we observed and what we expect if the genes are linked, is to be rejected.

References

Flagg, Raymond. 1988. Carolina Drosophila Manuel.

Morgan, Judith. Carter, M. Eloise Brown. Benjamin Cummings 2002. Investigating Biology: Fourth Edition.

http://media.nasaexplores.com/lessons/04-006/images/life-cycle.gif

Thompson, David. 2006. BIO 150L: Introduction to Biology 1 Laboratory Manual.

First off a special thanks goes out to Mike Combs and Main Street Baptist Church for helpin with Ally’s Dream! I don’t really know how to explain the rush of hearing those bubbles pop, but it was POP-O-LICIOUS! I don’t know if it was witnessing the smile on Ally’s face or what, but the enthusiasm from that night will reside in me forever! Another dream becomes a reality, another concept transcribes to life, and a contagious smile the only reward needed! Hopefully the video gets posted soon so you all can experience the thrill for yourself!

“I think you need to pick up a couple lit mags and see what people are writing, then mimick others until you find your own voice.”

This was advice given to me by a dude a while back. It’s stuck in my head ever since that day. For starters, he misspelled mimic! Why would anyone take advice from a self proclaimed expert when they are just as human as most the humans I know? This advice bothered me because it left me confused. The idea of mimicking others to find your own voice is weird. I get that evolutionary changes don’t occur overnight and that various small mutations accumulate, allowing a species to diverge, but why should one mimic another to find their own voice?
I know my voice, I hear it whenever I talk? I see it whenever I look in the mirror, and I feel it whenever a friend smiles! Besides, who would I mimic?

I think one of the issue with lots of people is that someone always assumes another person is going to fix the problem, do it different, do it better, do it bigger, etc. The issue with this is that that isn’t always the case. Look at the current state of the US economy. Who is responsible for this issue? How does a problem like that develop? Look at the issue’s concerning invasive species in Florida. How many people did it take assuming their small part wasn’t going to result in 150,000 pythons living in the state? How bad ass will the Florida ecosystem be in the future? The point of this is that when we choose to mimic other’s it’s usually out of respect or fear. When we mimic a hero, a very boomer mindset, it’s because we aspire to be like them. Cultural shift aside in the past decade, when we look towards others to find ourselves, we do so because we don’t fully understand who we are. Though it is totally cool to admire and aspire I ask how one finds their own voice when they are afraid to make their own path? Maybe my voice makes no sense, maybe my voice is unlike anything out there, maybe my voice uses made up words and does not recognize your imagery! What if my passion, my love, my lust, my drive has boundaries you can’t touch because your incapable of imagining the mechanical dragon wings that transform into rocket boosters, flying over a sea of crabs? What if we all made it a point to say what we wanted, regardless of the implications, and just lived life as a means to giggle instead of as a game to win? Cause at the end of the day, we all end up with the same outcome! No matter how well we play, how great we master, how powerful we become we end up as chaos!

I want to live in a world where love is safe, ignorance is frowned upon, fun is ok, and wantin to dance with @TheEllenShow isn’t crazy!

Cincinnati was ranked the craziest city in the country by some online poll, I heard that on the radio the other day. I just read it in the local newspaper too. This isn’t the good kind of crazy either, the survey took into account psychiatrist, stress, and drinking. We’ve already been voted the most dangerous city to live in. Cincinnati is also highly ranked in regards to cases of asthma and heart disease. You’d think Cincinnati rapper’s would be taken a little more serious by the city and the nation. Our hood is looking to be pretty damn tough! Hell, you know the streets are cursed when you can’t find a white man to dance with you, pretty much most days of the week

Then the poll goes and calls this dude from Jungle Jim’s our most colorful character? I resent that LOL! Maybe I need to step up my pumpkin suit campaign! I also started reading a book about generationme a.k.a. generation Y a.k.a. pretty much anyone who laughs at my videos! Apparently our whole generation believes we are entitled to chase our dreams. I think that this book is being a tad too harsh on us. We all have a right to chase our dreams. No matter what cultural influences shape our psyche, no matter the issue’s revolving around context and time frame, no matter the jibber jabber of coming to terms with reality, what purpose does life hold if we don’t make it a point to dream? What purpose does life hold if we’re not having fun, not messing with the normal way things are done, and don’t acknowledge crazy as the better alternative?

Outside of this nonsense let us talk about other types of nonsense!

1) Ever notice that the coolest girls to kiss are the ones who never admit to wanting to kiss you and actually tell you no, they hate you, while kissin on you?
2) Ever notice that really, really, really big people never seem to challenge you to foot races?
3) Why is it that all my black friends hate being called dark? Will I ever find a dark skinned chick happy that she is invisible on black-top?
4) Being invisible, though psychologically a trait desired by women according to many standardized evaluations, is dumb. Invisibility is totally a horrible super power. Think about it, crossing the street and some poor person wrecks their car into you. But the confused look on their dumbfounded face tells me I hope I witness an invisible person get hit by a car.
5) If all people were invisible, would we still fight or just assume we were alone?
6) Invisible movies would only be good for people with imaginations!
7) Invisible is apparently a tangent that I can’t get off! Invisible is like visible with an in, which is confusing to me because if your in and visible that means you’re recognized in some fashion. Why does taking two words that imply a composition of matter/status result in a meaning of nothing? Is this like a double negative?
9) Never piss off a moving house. That is not something many people will get, if you get it, then I hope you buy your home some rims!
10) If a committee of people get together to decide things are crazy, ever notice how a committee of people doing the crazy thing unites to fight for a cause?
11) Will we ever realize as a culture that things are only the things we say they are because people said?
12) Where do prions, viruses, bacteria, and countless other bi products of nitrogenous bases we’ve co-evolved with fall into Adam and Eve’s magical land?
13) Do people really believe plants can swim?
14) Why is football still called football? I hate Spanish lesson’s simply because of that; two sports, one name!
15) How is it that doing nice things for people isn’t always a “nice” thing to do?
16) When an ex girlfriend tag’s you in a conversation to trash talk you, is it normal to find that satisfying?
17) Why is it that most people seem interested in what you can do for them and not the other way around? At the very least what you can do together?
18) Is a PIMP still a PIMP if he has nobody to PIMP? Or better yet, is PIMPIN a state of mind, a way of life, or something you’re born with?
19) Cool is only cool when you have something to compare it to.
20) Simplicity comes out of ignorance or choice.

Today I volunteered at my mom’s school and taught first graders how to write! This time around I drew a picture of a giant alien on the white board. We then, as a class, wrote a story about Chipalow the alien! I love first graders!

I love how children have imaginations that are boundless! Children are not afraid to make ridiculous claims, are not afraid to say what’s on their minds, and at the end of the day giggle a ton. I admit the teacher wearin my t-rex chain was pretty sweet to!

BE CHILDISH! BE CREATIVE! HAVE FUN!

I’m so excited about the next random act’s of kindness. This coming 19th, a whole gym full of bubble wrap! Going to be great!

Oh I got some other surprises in the works for ya all! But I’ll keep you in suspense! This was much shorter than I expected, oh well, if your disappointed that means I LOVE YOU!

So today was a day of mixed feelings! The camera guy/producer/director who has been helping me for months work on the Ellen music video has pulled out of the project. Months I’ve spent working things around his schedule, pushed things back, and even turned away other camera guys because I put faith in this dude. This weekend I have a few people who were suppose to meet us and film a scene, including one with a backhoe! But I assume that is not going to happen and everything is going to be put back on hold.

If I’ve learned one thing, no matter how good someone is (this guy is good), never work with an ass hole. By that I mean individuals who are full of themselves, make it a point to put you down, and are totally self absorbed with themselves completely disregarding other people’s investment.

Lot’s of people have invested time and energy producing and mastering the Ellen song besides me. JD BONE from Hidden Studios, don’t think for a second mastering a song is an easy task. It took 6 plus hours to transfer the data due to Mac and PC issues alone. Then who knows how long he spent making everything crisp and nice. I know another few hours was invested re-recording. The dudes at Dubix, we spent 2 and a half hours trying to teach me how to rap on beat. That doesn’t even begin to scratch the surface of the time those guys put into making the “beat” and scheduling me studio time. Or let us not forget my amazing web guys, friends, family, etc. that have also invested energy and time into me talking about the song. Or my many fans who are waiting to hear and see it. Or Ellen who is following me on twitter because of it.

I put a lot of faith and trust in this man and took his word at face value. That is another lesson I guess I’m going to have to take from this. When doing any kind of production big ego’s override the justified thing to do. What makes me even more upset is that I spent 4 months waiting to get the mastered song done and truthfully should of already had the video shot. But my now ex camera guy made it a point he needed the final version to do what he needed to do.

People who are reading this, I advise you that if you do not want to do something, if you don’t want to work on a project, tell people up front. I have more respect for someone who says they don’t have the time, energy, or simply just don’t care than a person you invest in because they say they will do something resulting in and letting you and a ton of other people down. It’s OK, big surprises are in the works right now and this guy like all the other people who have flaked out along the way will miss out on the ride.

Disappointment or not, the show must go on! I guess you all are going to get a half arsed music video for now, unless I find another producer to work with. Preferably one who does not make it a point to tell me that my vision is wrong! With this said I’d like to tell all the people who haven’t flaked out THANKS! I love you all and together, one way or another, this is going to be well worth it. If nothing else, it’s been fun for me meeting and working with all of you!

I was the only dude sportin a pumpkin suit at the “Alice in Wonderland” movie! Totally fit the theme of the movie. I like the whole line about crazy people being the coolest! I paraphrase

What is the point of living a life without dreams? Where is it that we decide it’s better to end up being a tool in someones reality instead of pursuing our own passion? Where does the grand surreality of childhood vanish to?

When I was a little kid, I always wanted to grow up to be a paleontologist. Then I learned about digging holes and decided I wanted to be a dinosaur! Somewhere along the road the idea of being a real dinosaur faded. But why is that? If I walked around telling people my occupation was meat eater and roared at everyone who waved at me, people would want to put me in a mental institution! I don’t know about you, but that is not a fun place o be But regardless, it would be amazing! Seriously picture this, man waves and I go RAWR!!!!

I wanted to be a Tyrannosaurus Rex! I wanted to roar at and eat people I didn’t like! Never tell that to a stranger LOL! But imagine how cool life would be if we all had Jedi powers! How amazing would things be if we could really tame dragons and duke out our trivial wars with magic. Imagine for a second, being able to cast an army of a million winged spiders! A world without rules! Then some ass hole has to come along and say we need rules to protect us! People are going to hurt each other. What I want to know is where does that idea pop into our head? When I walked into a room full of little kids, even babies, some may have tempers, but I never see one worried about strangers. These ideas of danger and limitations, they are instilled in us someplace as we grow. Which is a shame. It’s a shame that as an invasive species capable of altering our environment we have not wrapped our heads around the delusion of control.

We all need dreams, for dreams give our lives a purpose, they give us a direction, for when we have no direction it doesn’t really matter which way we go. Then again, does it ever really matter at all? If we can’t verbalize with certainty why, then we might as well have fun! Delusional or not, fun is fun!