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- Using a schematic diagram, summarize the following steps in preparing competent cells for transformation: Inoculate a single colony of E. coli into 5 ml LB broth and incubate overnight at 37°C with moderate shaking (250 rpm). Add 200 μl of the culture into 50 ml LB broth and incubate overnight at 37°C with moderate shaking (250 rpm) to an OD600 = 1.3 to 1.5. Aliquot culture into five 15-ml pre-chilled, conical tubes. Leave tube on ice 5 to 10 min. Centrifuge cells 7 min at 1,600 × g (3,000 rpm), 4°C. Pour off supernatant and resuspend each pellet in 10 ml ice-cold CaCl2 solution (50 mM CaCl2), perform resuspension very gently, and keep on ice. Centrifuge cells 5 min at 1,100 × g (2,500 rpm), 4°C. Discard supernatant and resuspend each pellet in 10 ml ice-cold CaCl2 solution. Keep resuspended on ice for 30 min. Centrifuge cells 5 min at 1,100 × g, 4°C. Discard supernatant and resuspend each pellet in 10 ml ice-cold CaCl2 solution. Dispense cells (250 μl) into pre-chilled, sterile…General Electrophoresis Questions: 1. What makes macromolecules move through the gel in electrophoresis?2. What determines the speed at which macromolecules move through the gel in electrophoresis? In a single gel, why do some move faster than others?3. Why do we use different procedures for DNA and protein electrophoresis?"If you count 70 colonies from a urine culture obtained using 0.001ml calibrated loop, how many colony forming units (CFU) will be reported?" 70 700 7000 70000 700000
- Agarose Gel Electrophoresis Questions: 1. What determines how much agarose you should use in your gel? 2. Why do you see only DNA on the gel, and not protein? 3. How do you know which end of the gel to place the comb? 4. Suppose you turn on your power supply to run the gel and find that the milliamps reading is close to zero. What would you check?Copy and paste the link below and watch the video on Youtube https://www.youtube.com/watch?v=8RBs0Ghg_48 Answer the following Questions: 1. What are the chemicals and materials used in gel electrophoresis? 2. Draw a schematic diagram of a gel electrophoresis set-up 3. Describe the procedure in doing a gel electrophoresis experiment. Why is there a need for a leveling bubble/leveler? What is the use of the rubber dam? 4. What is the use of ethidium bromide and why must you wear gloves when you handle it? 5. What makes the DNA fragment move towards the positive plate? 6. What is the purpose of glycerol in the sample buffer? 7. What is the use of a DNA ladder? 8. What will happen when you increase the voltage of the set-up? 9. Can gel electrophoresis be used to separate amino acids? If so, how is it done?d/e/1FAIpQLSfTle9UfP15_VUqFI-ACEQd1XBykXv5Lr4dEMQbLJ1d6fCupw/viewform Students subjected three samples of five different molecules to gel electrophoresis as shown in Figure 1 A B C DE +2 3 Wells 4 8. Which of the following statements best explains the pattern seen on the * gel with regard to the size and charge of molecules A and B? 1 point molecules A and B are positively charged, and molecule A is smaller than molecule B. molecules A and B are positively charged, and molecule A is larger than molecule B. molecules A and B are negatively charged, and molecule A is smaller than molecule B. molecules A and B are negatively charged, and molecule A is larger than molecule B. Sign out
- What is the purpose of the blank test in the experiments?volume of the quasi-steady-state culture was V0= 500 L, and the nutrient solution containing glucose was added at a constant flow rate of F = 50 L/h.Data: X0 (at the beginning of feeding) = 20 g/L, S0 = 300 g/L, max = 0.2 h-1, KS = 0.5 g/L and Y x/s= 0.3 g/g a) Determine the volume of the culture at t = 10hb) Determine the concentration of glucose at t = 10 hc) determine the concentration and total mass of cells at t = 10 hd) If product is associated with growth with α = 1.5 and P0 = 0.1 g/L, determine the concentration of product at t = 10h. (answer:P = 67,55 g/L)MOLLISCH TEST You can use this as your reference : https://youtu.be/rKng5-ij6kQ
- How can you make a final dilution of 10-10 using 5 dilution tubes? Outline your steps.What side of the electrophoresis chamber (red or black) should the wells be on when the gel is placed in the chamber? What would happen if you placed the agarose gel with the wells on the opposite side of the chamber?Which of the following parameters should be altered to increase the amount of vector-insert constructs compared to other ligation products? vector:insert ratio o type of vector O incubation temperature O incubation time