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- You have just carried out a transformation using a plasmid (possessing a Amp-resistance gene) that was 0.001 ug/ul in concentration. You added 10 ul of this plasmid to 100 ul of a bacterial cell suspension. Then you added 250 ul of LB following heat shock. After plating 200 ul of this cell suspension with LB onto a LBA plate, you observe 8 colonies on this plate. What is the transformation efficiency of this plasmid (in colonies/ug of plasmid) based on these results? about 1400 O about 80O about 4000 O about 400O O about 8000 f6 米 IO fs fg f1o 19 08. O OOCalculate the transformation efficiency of an experiment conducted using 10µl of 0.005µg/µl plasmid, you plated 100µl out of a total volume of 500µl and the count of transformed colonies was 186.you have transformed E.coli cells with a plasmid containing the gene for the enzyme beta-lactamase you have used 3µl of a stock solution of 2.5ng/µl DNA. You transformed 150/µl of competent cells, using the calcium chloride method. you have added 600µl of luria broth to the cells and then plated out 200µl on triplicate plates. upon counting the colonies, you got the following results: plate 1: 120 cfu plate 2: 35 cfu plate 3: 95 cfu please answer the following question: 1. what new characteristic will the cells have ater transformation? 2. how are you going to test for the new characteristic? 3. calculate the transformation efficiency of the experiment?
- You are about to isolate a 3000 bp large plasmid from an E.coli culture. You know that the plasmid is present in 100 copies per E. coli cell. You aim to have a final plasmid concentration of 100 ng/µl in a total volume of 50 µl. Assuming the yield is 100 %, how many E. coli cells should the culture from which the plasmid is to be isolated at least containWhat is the role of the following in the alkaline plasmid screen? G buffer (Cell Suspension Solution) Denaturing Solution (Cell Lysis Solution) Neutralization Solution5 μL of plasmid DNA (pUC 19) was added to 50ul of CaCl2 competent E. coli that was then added 250 μL of SOC medium, 100 μL of this solution was plated onto a TSA + ampicillin plate. Can you please show me how I calculate the total number of successful transformants per mL of competent cells plated. The total number of colnies counted on plate was 70. Thank you
- The modifiedplasmid is reintroduced back into Rhizobium(step 4) and the genetically transformed bacteria are then selected based on the amp and lacZgenes present within the plasmid. The plasmid may or may not integrate the BBW resistancegene. The treated bacteria may or may not take up the modified plasmid. a) Complete the table below with a yes or no in each space stating whether you would expect these bacteria to grow or not. Type of treated bacteria culture plate(no amp) Culture plate treated with ampicillin Plasmid not taken up Plasmid taken up (WithoutBBW resistancegene) Plasmid taken up (WithBBW resistancegene) B) Outline how the genetically transformed bacteria containing the BBW resistance gene can beselected based on the amp and lacZgenes present within the plasmid.We use a NanoDrop spectrophotometer to analyze plasmid DNA obtained from miniprep. Whatinformation about the plasmid does the 260nm reading alone provide and why?You want to digest 1 µg of plasmid DNA in a final volume of 50 μL. Your solution containing plasmid has a concentration of 25 ng/μL. How many μL of your plasmid solution do you need to add to your reaction tube to digest your desired mass of plasmid?
- How does SYBR green work in DNA imaging and why does the uncut plasmid run faster than the cut plasmid? Please Explain. Thank youSince higher concentration colcemid will result in shorter chromosome, you want to change your protocol and reduce final concentration of colcemid in your 10 ml blood culture from 0.1ug/ml to 0.05ug/ml. how many ul colcemid stock solution with concentration10ug/ml needed to be added in 10ml blood culture?One of the frist steps in isolating plasmid DNA via mini-prep is to pellet the cells after O/N culture and then resuspend them in buffer P1. What's the point of pelleting the cells just to resuspend them again? To concentrate the cells P1 is a buffer, preventing small changes in pH O This step is not absolutely necessary P1 begins the lysis process D Question 2 What would happen if we didn't centrifuge the tube containing lysed cells after adding neutralization buffer and instead just added right to the column? O We could fihish the prep but we would have protein/RNA contaminants at the end O The precipitate is less dense than water so it shouldn't get in the way since it floats on top O The precipitate would obstruct the column and our prep is ruined O Our yield would be reduced, but we'd get something