Biology
5th Edition
ISBN: 9781260487947
Author: BROOKER
Publisher: MCG
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Chapter 21.1, Problem 1CC
Summary Introduction
To determine: The purpose of the lacZ gene in the vector in the cloning experiment.
Introduction: Genomics contains an important technique by which multiple and exact copies of a gene can be produced. This technique is known as gene cloning. This method requires certain agents such as plasmids and viruses for the successful production of multiple genes from a specific gene. These are known as vectors. Vectors carry the gene of interest in the host cell and thus, help in gene cloning.
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Preparing plasmid (double-stranded, circular) DNA for sequencing involves annealing a complementary, short, single stranded oligonucleotide DNA primer to one strand of the plasmid template. This is routinely accomplished by heating the plasmid DNA and primer to 90 °C and then slowly bringing the temperature down to 25 °C. Why does this protocol work?
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Assume you were given competent cells of known transformation efficiency (TE). Assume TE= 1×10[6] (note 10[6] means 10 to the power of 6). You want to have about 1000 colonies on the P-200 plate. How many nanograms of plasmid should you use in the transtormation reaction?
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Chapter 21 Solutions
Biology
Ch. 21.1 - Prob. 1CSCh. 21.1 - Prob. 1CCCh. 21.1 - Prob. 2CCCh. 21.1 - Prob. 3CCCh. 21.2 - Prob. 1CCCh. 21.2 - Prob. 2CCCh. 21.2 - Prob. 3CCCh. 21.3 - Prob. 1EQCh. 21.3 - Prob. 2EQCh. 21.3 - Prob. 3EQ
Ch. 21.4 - Prob. 1CCCh. 21.4 - Prob. 1CSCh. 21.5 - Prob. 1CSCh. 21.5 - Repetitive Sequences and Transposable Elements...Ch. 21 - Prob. 1TYCh. 21 - DNA ligase is needed in a cloning experiment a. to...Ch. 21 - Prob. 3TYCh. 21 - Why is Taq polymerase used in PCR rather than...Ch. 21 - Lets suppose you want to clone a gene that has...Ch. 21 - In the CRISPR-Cas technology for editing genes,...Ch. 21 - Prob. 7TYCh. 21 - The enzyme that helps short segments of DNA move...Ch. 21 - Prob. 9TYCh. 21 - Which of the following was not a goal of the Human...Ch. 21 - Prob. 1CQCh. 21 - Briefly describe whether or not each of the...Ch. 21 - Prob. 3CQCh. 21 - Identify and discuss three important advances that...Ch. 21 - Prob. 2COQ
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- A. Please briefly explain how Polymerase Chain Reaction works to amplify DNA. B. Please briefly explain what gel electrophoresis is and how it works to separate a mixed sample of macromolecules like DNA. C. Briefly describe what a plasmid is, and how it can be used to transform bacteria like E. coli.arrow_forwardPreforming a "blue-white screen" 3) Would bacteria that have taken up a plasmid into which a DNA fragment has been inserted, form a blue colony or a white colony when grown on this medium? Briefly explain why these bacteria would form a colony of the color you chose.arrow_forwardAttached is an image of EcoRI-digested plasmid samples (pBSK and pEMBL1.9); explain why the bands are found at the top, the purity of the results, and how we couple improve our results.arrow_forward
- 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 sitearrow_forwardGive typed full explanation you made a recombinant plasmid (it is circular) that contains the sequence 5' AAGCTT in the middle of the inserted or cloned gene. that is the only place where this sequence occurs. the entire plasmif does not have 5' GAATTC. When putting this plasmid into E. coli (transformstion) what will happen? 1. plasmid will cut into several fragments 2. nothing the plasmif with remain circular 3. plasmid will be cut once making a linear dna fragment 4. will be cut once but will becomr two linear fragmentsarrow_forwardA.How could endonucleases interfere with the transformation procedure? B. Does supercoiled or nicked plasmid get transformed more efficiently? Why?arrow_forward
- 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.arrow_forwardDNA molecular with complementary sticky ends associate by (a) covalent bonds (b) hydrogen bonds (c) ionic bonds (d) disulfide bonds (e) phosphodiester linkagesarrow_forwardn gel electrophoresis of DNA, the different bands in the final gel form because the DNA molecules _______. a. are from different organisms b. have different lengths c. have different nucleotide compositions d. have different genesarrow_forward
- Consider the image below. This image shows plasmid DNA isolated through exactly the same method that you used. The lanes that were untreated (TE) contain plasmid DNA, but also large amounts of smaller nucleic acids that are predicted to be RNA. What is the experimental evidence from the gel itself that these smaller molecules are RNA? (Hint: look at the treatments performed in each lane)arrow_forwardstate two ways one can extract an insert from a plasmidarrow_forwardMany studies have linked plasmids to bacterial resistance to antibiotics. Closured circular segments of double-stranded DNA, plasmids have several restriction enzyme cut sites along with genes that give the cell advantageous characteristics. Do you believe that fighting widespread antibiotic resistance may be accomplished by focusing on these specific DNA segments? In one or two paragraphs, formulate a well-supported opinion. Can you please provide a citation as wellarrow_forward
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