9. Here is a restriction map for a bacterial plasmid showing the cleavage sites for two different restriction enzymes EcoRI and Bam H1. EcoR1 700 bp EcoR1 2000 bp 1200 bp BamH1 Match the banding pattern in each lane of the agarose gel below with one of the treatments listed. Note: Not all treatments listed are run on the gel. Plasmid treatments: A. BamHI+EcoR1 (no RNAse) B. BamHI only (no RNAse) Gel lanes: 1. DNA Ladder = Mol. Wt. Standard (sizes indicated) bp 5000 4000 3000 2000 1650 1000 850 650 500 C. BamH1 + EcoR1 + RNAse D. RNAse only 2. E. BamH1 + RNAse F. EcoR1+ RNAse 3. G. EcoR1 only (no RNase) 4. 1 2 3 4 5 6 5. 6. -

9. Here is a restriction map for a bacterial plasmid showing the cleavage sites for two different restriction enzymes EcoRI and Bam H1. EcoR1 700 bp EcoR1 2000 bp 1200 bp BamH1 Match the banding pattern in each lane of the agarose gel below with one of the treatments listed. Note: Not all treatments listed are run on the gel. Plasmid treatments: A. BamHI+EcoR1 (no RNAse) B. BamHI only (no RNAse) Gel lanes: 1. DNA Ladder = Mol. Wt. Standard (sizes indicated) bp 5000 4000 3000 2000 1650 1000 850 650 500 C. BamH1 + EcoR1 + RNAse D. RNAse only 2. E. BamH1 + RNAse F. EcoR1+ RNAse 3. G. EcoR1 only (no RNase) 4. 1 2 3 4 5 6 5. 6. -

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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Which of the following is not correct for bacterial conjugation? a. Both Hfr and F+ bacteria have the ability to code for a sex pilus. b. After an F- cell has conjugated with an F+, its plasmid holds the F+ factor. c. The recipient cell is Hfr following conjugation. d. In an HfrF- conjugation, DNA of the main chromosomemoves to a recipient cell. e. Genes on the F factor encode proteins of the sex pilus.
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
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. Given the following restriction endonucleases and the sequences of their corresponding restriction sites, which would be LEAST useful for cutting the plasmid and the foreign DNA to be inserted? [The arrows indicate where cuts are made.] (see img) a. EcoRI b. HindIII c. HaeIII d. BamHI 2. EcoRI and HindIII are two different restriction enzymes. If the DNA from two different sources were cut with either EcoRI or HindIII, which of the cut DNA fragments would NOT join together easily so that they could be sealed with ligase? a. E. coli DNA cut with EcoRI and human DNA cut with HindIII b. E. coli DNA cut with HindIII and mouse DNA cut with HindIII c. human DNA cut with EcoRI and chimpanzee DNA cut with EcoRI d. mouse liver DNA cut with HindIII and mouse kidney DNA cut with HindIII
1. Below is an illustration of two plasmids: the incomplete pKAN-R which contains the gene of interest (rfp), and the good plasmid vector pARA which contains the ampR gene for ampicillin resistance, the araC activator gene and the ori region. Based on this, how are you going assemble to create the desired recombinant plasmid containing all the necessary components? (Keywords: Identify correct restriction enzymes to use to create specific sticky end sequences that will only assemble with the final vector in one orientation)
83. A plasmid is cut with the restriction enzyme BamH1 giving fragments of 3000 and 1000 bp as identifiedby gel electrophoresis and ethidium bromide staining. In a seperate restriction digest the enzyme EcoR1gives fragments of 1000 and 1500 bp that are apparent on the agarose gel. What is the most likely size ofthe plasmid in bp? Explain why it's 4000
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)
3A. Following transformation with the CRISPR plasmid, the yeast will be plated on SD-URA plates. Why is SD-URA media used? Name the relevant marker gene in the pCRCT CRISPR plasmid. 3B. What would you expect to see if you plated on YED accidentally? Will most of the yeast be red or white? Why? 3C. The complete genotype of the host yeast strain is MATa ade2 his3 leu2 met15 ura3. Name one advantage of using a yeast strain with auxotrophies for several genes (i.e. his3, leu2 and ura3). 3D. After successful CRIPSR editing, the yeast can later be “cured” of the recombinant CRISPR plasmid – that is, the plasmid is lost but the CRISPR edit is stably inherited. Write the new genotype of the yeast-based on the ADE6 gene.
Hey, I need help with this please: Plasmid pRIT450 is 7.0 kb in length and has single PstI, EcoRI, and BamHI sites. I have cut the plasmid with PstI and inserted a 4.0 kb fragment into the site. From the data below, I need to Produce a "restriction digest map" of the resulting plasmid. Please draw the "restriction digest map" with the given data and also explain the sequence of steps you took to construct the digest map so I can understand it thoroughly.
You utilised two plasmids in this practical, pOTC and pOTC-Δ. Plasmids are often represented using plasmid maps like the one below. This map shows the positions of recognition sites for a number of restriction enzymes Using the plasmid map of pBCH2.0 provided above, predict how many DNA fragments would be formed if this plasmid was digested with restriction enzymes EcoRI and PvuII.
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. Match the features of plasmids below with their meaning : Origin of replication LacZ gene antibiotic resistance gene Multiple cloning site A.required for maintenance of the plasmid in E. coli cells used to select for coli that contain our plasmid B. Required for replication of plasmid DNA inside an E. coli cell C. An engineered region of the plasmid that contains many unique restriction enzyme recognition Sites D. Allows for screening for plasmids with an insert in the multiple cloning site