Becker's World of the Cell (9th Edition)
9th Edition
ISBN: 9780321934925
Author: Jeff Hardin, Gregory Paul Bertoni
Publisher: PEARSON
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Chapter 18, Problem 1Q
Of these three techniques, which one provides the most specific information about where a protein binds to DNA? Which one(s) require(s) specific knowledge about the DNA sequence bound by the protein before performing the procedure?
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Both protein and DNA are run together in an isoelectric focusing (IEF) electrophoresis using the immobilised pH gradient (IPG) strip with pH range of 4-7. After the electrophoresis and staining, only ONE band is observed on the middle of the IPG strip. The band is a protein band. Briefly explain why only the protein band and NOT the DNA band appear on the IPG strip.
You have an extract of BL21(DE3) cells containing a soluble, 75 kDa recombinant protein that has a histidine tail at its C-terminus. This extract is still in raw form and you need to purify this protein without losing its solubility. Check the option below that represents the best purification methodology in the order of execution. *
a) (1) isoelectric precipitation; (2) microfiltration; (3) affinity chromatography; (4) anion exchange chromatography b) (1) centrifugation; (2) microfiltration; (3) isoelectric precipitation; (4) affinity chromatography c) (1) centrifugation; (2) microfiltration; (3) affinity chromatography; (4) ultrafiltration
d) (1) microfiltration; (2) jumping out; (3) centrifugation; (4) affinity chromatography
Describe the function of the following reagents used in the DNA extraction procedure?a) Proteinase K b) 5M Nacl c) Isopropanol d) 1X TE Buffer
Chapter 18 Solutions
Becker's World of the Cell (9th Edition)
Ch. 18 - Suppose a triplet on the template strand of a...Ch. 18 - Of these three techniques, which one provides the...Ch. 18 - Compare and contrast bacterial and eukaryotic...Ch. 18 - The autoimmune disease systemic lupus...Ch. 18 - QUANTITATIVE Triplets or Sextuplets? In his Nobel...Ch. 18 - The Genetic Code in a T-Even Phage. A portion of a...Ch. 18 - Frameshift Mutations. Each of the mutants listed...Ch. 18 - Prob. 18.4PSCh. 18 - Locating Promoters. The following table provides...Ch. 18 - Prob. 18.6PS
Ch. 18 - Starting Up. Refer to Figure 18-30, which depicts...Ch. 18 - RNA Processing. The three major classes of RNA...Ch. 18 - Prob. 18.9PSCh. 18 - Antibiotic Inhibitors of Transcription. Rifamycin...Ch. 18 - Prob. 18.11PSCh. 18 - Cloning Conundrum. Using established recombinant...Ch. 18 - Nucleoli. Indicate whether each of the following...
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- In standard agarose gels used for the analysis of DNA, linear DNA fragments are separated on the basis of their... a) Size b) Charge c) Buoyancy d) Color e) Smellarrow_forward1) The Qiagen DNeasy DNA extraction kit is used to extract DNA from a liver cell line for subsequent molecular experiments. (i) (ii) Explain the principle behind this kit in DNA extraction. After the DNA is obtained, the purity of DNA should be determined. Discuss the steps for checking DNA purity using a spectrophotometer.arrow_forwardAgarose gel electrophoresis is a technique used to separate DNA fragments on an agarose matrix. The DNA is then viewed by staining or under UV light. (i) What is the direction of DNA migration in electric field on the agarose gel? · . (ii) Explain the principle for the migration of DNA molecules in an electric field. (iii) DNA loading dye is used to prepare DNA markers and samples for loading on the agarose gel. State the function of bromophenol blue in the loading dye.arrow_forward
- Explain why DNA fragments migrate in a gel electrophoresis. Which fragments migrate farthest: large or small?arrow_forwardDescribe the common strategy (steps) for protein sequencing, starting with a biological sample containing many cell and biochemical substances. How prevalent are disulfide links in proteins? Why do the disulfide links need to be broken prior to sequencing? How can they be chemically broken?arrow_forwardThe oligonucleotide d-ATGCCTGACT was subjected to sequencing by Sanger’s dideoxy method, and the products were analyzed by electrophoresis on a polyacrylamide. Draw a diagram of the gel banding pattern obtained.arrow_forward
- TRUE OR FALSE a) The long solenoid structure of the chromatin material binds to a protein scaffold through the SARs or MARs in the DNA. b) There can be a quantitative determination of the degree of supercoiling in a DNA sample.arrow_forwardCystic fibrosis (CF) is an inherited disorder caused by different types of mutations, many of which prevent ions from moving across cell membranes. Normally there are channel proteins that allow passage of the ions, but in patients with one kind of CF these proteins seem odd. Closer examination shows that these proteins display the correct amino acid sequence. However, they fail to do their job. A) Given that the primary structure of the protein is correct, what can you infer about the DNA sequence for the gene coding this protein on this patient, is there a mutation? Explain. B) Why is the primary structure insufficient to guarantee the proper function of the protein?arrow_forwardDideoxysequencing relies on which one of the following choices: A):Random stopping of one of many sequence reactions by insertion of a dideoxynucleotide B):Random stopping of one of many sequence reactions by insertion of a deoxynucleotide C):Random initiation of one of many sequence reactions by insertion of a dideoxynucleotide D):Having at least two dideoxynucleotides per chain E):The inability of DNA polymerase to incorporate a dideoxynucleotidearrow_forward
- During agarose gel electrophoresis, why does DNA move through the gel when electric current is applied? because DNA is negatively charged because a charged chemical from the loading buffer is bound to the DNA because DNA is positively charged because DNA absorbs electricityarrow_forwarda) Explain the effect of the guanine:cytosine ratio on melting temperature of DNA. b) The Hershey-Chase experiment proved that DNA is the genetic material and not protein. Explain in detatil how this experiment was conducted.arrow_forwardwhat type of gel(in terms of material) can be more suitable for the electrophoresis of 500-1000bp DNA fragment and why?arrow_forward
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