Genetically modified organismsBacterial transformation and protein purificationCrossing Over in Flies10. (0.5 points) For each of the three sets of data, calculate the % crossover between the genes based on the F2 phenotype counts. For this, assume that crossing over happens at equal rates in males and females. Show your work.They should show their work, but the formula is f(crossing over) = 1-(2(√ f(aabb))), which was covered in the lecture. The “f(aabb)” is the % (in decimal form) of the double recessive phenotype (either white or sepia&ebony) so they’ll have to calculate that first from the fly counts.11. (0.5 points) What is the distance between each pair of traits in map units to the nearest tenth of a M.U.? Are there any values that are problematic? Explain why if yes or why not if no.M.U. is equal to the % recombination, so in order from above the MUs are 35.05, 26.46 and 30.25 M.U., respectively.The problematic value is for st & bw. These two genes should be unlinked traits (they are on different chromosomes) so we expect them to calculate to a M.U. of at (or close to) 50 M.U. (since 50% rates are equivalent to unlinked).Name: ___________________________ Lab room, day, time: __________________TA name: _______________Genetically modified organismsYou will work with your own GMO data.1. The procedure you followed for testing for GMO foods makes heavy use of controls. In fact, only one PCR reaction was not a control. It also has the interesting outcome that DNA or primers may be used in common between both positive and negative controls, making the identification of each type tricky.a. (1 point) Identify all of the positive control PCR reactions. Briefly defend your choices.The key difference between positive and negative controls is that positive controls should give you a measurable response, while negative controls should have the absence of the measurable feature. So any reaction that was supposed to give a band would be a positive control, regardless of the specific combinations of templates and primers. They are: GMO+ template DNA with GMO-specific primers.GMO+ template DNA with plant-specific primers.Test food DNA with plant-specific primers.Non-GMO food DNA with plant-specific primers.b. (1 points) Identify all of the negative control PCR reactions. Briefly defend your choices.Same reasoning as above, except the negative controls are those reactions that should notproduce a band. The only one is:Non-GMO food DNA with GMO-specific primers.2. (1 point) What was your tested food? Was it genetically modified? If yes or no, explain with regards to the controls how you reached that conclusion. If your results could not be interpreted accurately, explain why not.Answers will vary. The soybean tested positive for GMO last semester and the cornmeal is a known one as well, but the others won’t be known until they are actually tested.Reasoning for a yes: There were bands for all of the positive controls and no band for thenegative control. There was a band in the test food DNA with GMO-specific primers and it corresponded to the band in the GMO+ template DNA with the GMO-specific primers positive control.Reasoning for a no: There were bands for all of the positive controls and no band for the negative control. There was not a band for the test food sample with the GMO-specific primers (meaning that in general the banding pattern should match that of the non-GMOPCRs).Couldn’t interpret: Answers will vary depending on what specifically went wrong. A missing band in a positive control indicates that something did not work (an extreme would be no bands in any samples). Having a band in the negative control might indicatecross-contamination, which would shed doubt on a band being seen in the test food with GMO. Bands of different sizes than expected could also indicate problems with the PCR.3. (1 point) Animals can be genetically modified as well. What components of the PCRs need to be changed in order to identify genetically modified meats or milk? Generally explain why those changes need to be made.Obviously the chloroplast primers wouldn’t detect anything from animals, so there would need to be a different positive control for something found in animals. Students could give examples (maybe something like myosin II for meats), but “animal-specific” is fine. Additionally, the GMO-specific primers are against sequences for a promoter from a plant virus and a terminator from a plant bacterium. These GMO-specific primers would need to be changed to whatever was commonly being used to transform animals (so maybe sequences from animal viruses). The template DNA would be different since it would be from the tested animal products. That is a bit of a no-brainer so they don’t have to specify it. However, they should mention the change to the GMO-positive template DNA since the primers would be different.Bacterial transformation and protein purificationYou will work with your own (or possibly borrowed) observations this week.Report the observations of growth and fluorescence for the plates you examined.PlatesGrowth observationsColony Color underUV light-pGL O/LB-pGL O/LB /amp+pGL O/LB /amp+pGL O/LB /amp/araThe above shouldn’t be graded, it’s just a check for you to be able to appropriately grade question 5.4. (1 point) The –pGLO plates are controls. Identify if each is a positive or negative control and explain why it is used (that is, what information does it supply).The –pGLO/LB plate is a positive control. Since it only contains nutrient medium, any bacterial cell should grow. This lets you know if your bacteria are generally viable (sincea lack of growth on this plate would indicate a fundamental problem with the bacteria).(In the generic terms, this protects against false negatives.)The –pGLO/LB/amp plate is a negative control (since we don’t expect to see growth). Since the bacteria should naturally not be antibiotic resistance, this shows us if that is actually true. Any growth on this plate would indicate a problem with that (and in more detail might inform us of a background rate of resistance, but the students don’t have to mention that.(In the generic sense, this protects against false positives.5. (0.5 points) Did the observed patterns match your expectations based on an understanding of the bacteria and plasmid? Why or why not?Answers will vary based on what results they actually saw. Plain LB plate should show lawn growth, no glow,