10. After transformation, you are interested in your plasmid efficiency. You saw 90 colonies after overnight incubation on a selective media. You remembered that you used 0.1 microgram of plasmid DNA and that you used a mere 100 microliter to plate colonies from a total volume of 1 mL cell. Calculate the transformation efficiency.

10. After transformation, you are interested in your plasmid efficiency. You saw 90 colonies after overnight incubation on a selective media. You remembered that you used 0.1 microgram of plasmid DNA and that you used a mere 100 microliter to plate colonies from a total volume of 1 mL cell. Calculate the transformation efficiency.

Biology: The Dynamic Science (MindTap Course List)

4th Edition

ISBN:9781305389892

Author:Peter J. Russell, Paul E. Hertz, Beverly McMillan

Publisher:Peter J. Russell, Paul E. Hertz, Beverly McMillan

Chapter18: Dna Technologies: Making And Using Genetically Altered Organisms, And Other Applications

Section: Chapter Questions

Problem 3TYK: Why are antibiotic resistance markers such as ampR important components of bacterial plasmid cloning...

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Similar questions

What information and materials are needed to amplify region of DNA using PCR?
1. You are having problems getting good transfection efficiency when you put your plasmid in e. coli. You want to freeze down a stock of e. coli that is more than 99% transfected. What modifications can you make to your plasmid and what steps can you take to ensure that 99% of your e. coli are transfected?
3. Another common electrophoresis buffer is TBE. How does this buffer differ from TAE? (plasmid purification experiment)
4. What is the name of the process by which bacteria pick up a different organism’s geneticmaterial?5. Genetically, how does the original bacterial DNA (plasmid) differ from the final bacterial DNAmolecule? (Do not say, “It is longer.”)6. Tetracycline is an antibiotic that is prescribed to kill bacterial infections. The transformedbacterial cell that you created has the tetracycline resistant gene in it. If this cell is placed on agrowth medium with tetracycline, will the bacteria grow or die? Explain.
2. Why do you increase the pH to denature the plasmid and chromosomal DNA during alkaline lysis rather than usinghigh temperatures, which would also denature DNA?
1. If you do an absorbance reading after plasmid purification and get a A260/A280 of less than 1.8, how could you further purify the sample to get rid of the protein contamination? Is it always necessary to have completely pure DNA? What are some cases where it would or would not be?
1. Write if the statement is true and modify the statement with the correct answer if its false a.White colonies represent those bacteria that do not express the genes for beta-galactosidase because restriction enzymes have cut the gene and ligase have not repaired the break b.Antibiotic resistance genes in bacterial transformation are used as marker genes because of ease of detection of transformants. c. Bacterial transformation simply means the insertion of a recombinant plasmid into a bacterial cell.
1 (a) What do ApR, TcR, and ori on the pBR322 map represent and discuss there individual functions? (b)Does the undigested plasmid show more than a single band when electrophoresed? Why? (c)What other kinds of molecules, in addition to plasmid DNA would you expect to find in a sample of plasmid DNA extraction? (This is not a graded assignment)
9. Estimate the sizes of the bands, in each of the lanes for the gel attached below. Then, for each lane, match the fragments created with the set of restriction enzymes used in the digestion of 640 bp DNA.
2. You are cloning the genome of a new DNA virus into pUC18. You plate out your transformants on ampicillin plates con-taining X-gal and pick one blue colony and one white colony. When you check the size of the inserts in each plasmid (blueand white), you are surprised to fi nd that the plasmid fromthe blue colony contains a very small insert of approximately60 bp, while the plasmid from the white colony does notappear to contain any insert at all. Explain these results.
1. Write if the statement is true and modify the statement with the correct answer if its false a.Blue colonies in the white-blue screening provide sufficient information on the transfer of recombinant DNA in bacterial cells. b.In blue-white screening, it is only the blue colonies that have survived the effect of antibiotics in the media. c.Bacterial transformation simply means the insertion of a recombinant plasmid into a bacterial cell. d.Competent cells are those bacterial cells that can accept the entry or re-entry of plasmids.
1. Briefly explain why the total size of the pMBBS plasmid in the Restriction Enzymes practical is 3000bp (base pairs) 2. Briefly explain why the cut sites on the pMBBS plasmid in each Restriction Enzymes (EcoRI, BamHI, and XhoI) just 1 each 3. Briefly explain what led to the 5 fragments formed which linked to the 500bp, 1000bp, 1500bp, 2000bp, 2500bp sizes