I need help 1)You are working with dye samples. In electrophoresis, researchers will commonly add a loading dye to a DNA sample when running a gel; WHY? 2.) What two factors of the samples control the length of distance the dye will migrate? 3.) What applied force pushes the dyes through the gel? 4.) Agarose electrophoresis is commonly used to separate molecules of DNA. Consider how DNA molecules with molecular weights of 450, 2000, 600, 5000, and 1000 daltons would separate; list them in the order they would resolve on the gel if they were loaded from the (-) anode and run towards the (+) cathode? 5.) Examining your final gel, did each sample form an individual band? If any of the samples formed more than one band, how did that reflect the sample's overall color? Remember the color wheel and how primary colors mix together to form secondary colors. 6.) Why is DNA and RNA typically loaded into wells placed at the negative end of the electrophoretic kit? 8.) Do you think the unknown sample represents a new mane color for My Little Ponies? If you answer 'yes', think of a name for the new My Little Pony character and write it here. Determine what factors are contributing to its unique coloration? 9.) Multimillionaire Cardi B is the wealthiest person to come out of the Bronx in recent history (sorry Ice!). Her favorite hair dye is IZZY BLUE! In fact, she had a researcher produce a series of bacteria that have the genes genetically engineered into them to create the blue dye so she can mass-produce it and market it as Cardi's Blue! Unfortunately, the wildtype strain developed multiple LOF mutations and no longer produce the blue product. The same researcher who created them suggests Cardi try interacting them with one another to see if any of the mutant strains can 'rescue the wildtype'. Cardi is not familiar with the terminology so she needs your help in interpreting the complement chart outcome. a. How many mutant genes are indicated in the chart? b. Which mutants does Cardi need to interact in order to produce Cardi Blue again? Please show how you figured out your solution. Figure 2. Cardi says 'Don't be letting me down! I ain't here for it!' X B1- B2- B3- B4- B5- B6- B7- B1- 0 B2- 0 0 B3- X X 0 B4- 0 0 X 0 B5- X X 0 X 0 B6- 0 0 X 0 X 0 B7- X X 0 X 0 X 0 Figure 3. Cardi Blue bacterial mutant outcome following a complementation test. 10.) A genetic BRONY pathway has been elucidated that results in My Little Ponies with black manes. Researchers suggest three genes are in the pathway, Bro1+, Bro2+, and Bro3+. Each gene produces a pigment with the color black resulting from a fully functional wildtype. Three LOF bro' mutants have been discovered that affect the outcome of the pathway. That each mutant represents a different gene is shown by the complementation chart below. When each mutant is interacted with one another, the wt pigment is restored. 10A.) How many genes does the complementation chart outcome suggest?___________ X Bro1- Bro2- Bro3- Color Bro1- 0 + + black Bro2- + 0 + black Bro3- + + 0 black When the LOF mutants are kept isolated and not complemented, the colors they produce are different, as shown in the results below. This gives an indication of what color pigment is produced by each gene. Organism Genotype Color ofmane Wildtype Bro1+ ; Bro2+ ; Bro3+ black Mutant 1- Bro1- ; Bro2+ ; Bro3+ purple Mutant 2- Bro1+ ; Bro2- ; Bro3+ orange Mutant 3- Bro1+ ; Bro2+ ; Bro3- green That each wt gene produces a different color is shown by the following feeding chart where each LOF mutant was 'fed' the pigment it cannot produce. Notice the difference in layout between a complementation chart and a feeding chart. Mutant/Supplied With Yellow Color Red Color Blue color Bro1- black purple purple Bro2- orange orange black Bro3- green black green 10B.) Based on the outcome of the three charts, determine what color is produced by each of the three genes, Bro1+, Bro2+, and Bro3+. Refer to the following color wheel on color theory if you are having trouble answering this question. Bro1+ = _____________________ pigment Bro2+ = _____________________pigment Bro3+ = _____________________pigment 10C.) Considering the outcome, describe this BRONY pathway using one of the following words: pleiotropic, polygenic, epistatic, penetrance, or expressivity. Explain your choice. 2 EC points. In a few sentences, describe how you could determine the size of molecules using the process of gel electrophoresis. Extra credit questions (2 points each). Q1. Bad Bunny gets sidetracked putting a new coat of polish on his nails while running DNA samples out on a gel and mistakenly lets it run for 100 minutes instead of 15 minutes! When he retrieves the gel, there are no bands on it?! Dios Mio! Where are the samples? A1. Q2. Dr. Duh is planning on separating samples of Titin protein molecules, the largest protein found in humans, using electrophoresis. Would he need an agarose gel with a higher concentration of agarose or a lower concentration? Explain your response. A2