Biochemistry: Concepts and Connections (2nd Edition)
2nd Edition
ISBN: 9780134641621
Author: Dean R. Appling, Spencer J. Anthony-Cahill, Christopher K. Mathews
Publisher: PEARSON
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Chapter 4, Problem 8P
Refer to Figure 4.15, which presents the Meselson-Stahi experiment. DNA molecules can be denatured by high pH, as well as by heat. Suppose that the CsCl gradient centrifugations were run at pH 12, conditions under which DNA strands separate. Sketch the gradient profiles expected for each of the four samples depicted in the figure.
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Answer part C of the following:
A) E. coli DNA and binturong DNA are both 50% G-C. If you randomly shear E. coli DNA into 1000 bp fragments and put it through density gradient equilibrium centrifugation, you will find that all the DNA bands at the same place in the gradient, and if you graph the distribution of DNA fragments in the gradient you will get a single peak (see below). If you perform the same experiment with binturong DNA, you will find that a small fraction of the DNA fragments band separately in the gradient (at a different density) and give rise to a small "satellite" peak on a graph of the distribution of DNA fragments in the gradient (see below). Why do these two DNA samples give different results, when they're both 50% G-C? See graph B.
B) If you denatured the random 1000 bp fragments of binturong DNA that you produced in question 1a by heating them to 95ºC, and then cooled them down to 60ºC and allowed them to reanneal, you would find that approximately 15% of the…
It is desired to isolate genomic DNA from liquid culture of S. cerevisiae yeast. A commercial kit will be used to isolate genomic DNA from this liquid culture. Answer the following questions to understand the strategy used by commercial kits for genomic DNA isolation.
a) List all the steps from cell pellet preparation to DNA elution.
b) With which feature can the membrane in the column that comes with the commercial kit bind DNA?
c) Which component in the kit would you use to recover the DNA from the membrane of the column to which the DNA was attached?
The following DNAs were incubated with E. coli DNA Polymerase I and dNTPs. Write the products expected in each reaction. Indicate the activity used in each reaction.
Chapter 4 Solutions
Biochemistry: Concepts and Connections (2nd Edition)
Ch. 4 - Prob. 1PCh. 4 - What is the difference between a nucleoside...Ch. 4 - pppApCpCpupApGpApu-OH a. Using the straight-chain...Ch. 4 - Shown is a representation of a molecule being...Ch. 4 - Base analysis of DNA from maize (com) shows it to...Ch. 4 - Using the pKa data in Table 4.1 and the...Ch. 4 - For some DNAs, it is possible to separate the two...Ch. 4 - Refer to Figure 4.15, which presents the...Ch. 4 - Suppose that you centrifuged a transfer RNA...Ch. 4 - Predict the structure of a cruciform that could be...
Ch. 4 - DNA from a newly discovered virus was purified,...Ch. 4 - Would you expect Neurospora crassa DNA to have a...Ch. 4 - A circular double-stranded DNA molecule contains...Ch. 4 - The gel electrophoresis pattern in Figure 4.23 was...Ch. 4 - 15. DNA polymerase requires both a template, to be...Ch. 4 - Prob. 16PCh. 4 - Prob. 17PCh. 4 - a. What two enthalpic factors stabilize DNA in...Ch. 4 - 19.
a. The plasmid pBR322 (4362 base pairs) was...Ch. 4 - Prob. 20PCh. 4 - What DNA sequence feature is required for a...
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Need a deep-dive on the concept behind this application? Look no further. Learn more about this topic, biochemistry and related others by exploring similar questions and additional content below.Similar questions
- Consider the experiment conducted by Meselson and Stahl in which they used 14N and 15N in cultures of E. coli and equilibrium density gradient centrifugation. Draw pictures to represent the bands produced by bacterial DNA in the centrifuge tube before the switch to medium containing 14N and after one, two, and three rounds of replication in that medium. Use separate sets of drawings to show the bands that would appear if replication were (a) semiconservative; (b) conservative; (c) dispersive.arrow_forwardExplain the rationale behind the separation of different cellular components using centrifuge. Explain the importance of adding isopropyl as the final solution to isolate the concentrated DNA. What is the chemistry basis of this step?arrow_forwardExplain the principles behind DNA isolation using silica membrane spin column from Qiagen. 2 After DNA isolation using Qiagen extraction kit, the final flow through was subjected to spectrophotometric measurements. Explain the situation if the absorbances were found to be: i) The absorbance at 260 nm is higher than absorbance at 280 nm? ii) The absorbance at 230 is higher than absorbance at 260 nm?arrow_forward
- PCR is a molecular biology technique where template DNA is amplified using a primer and oligonucleotides. The reaction is catalyzed by a thermostable DNA polymerase and in a particular reaction, the template strands are denatured at 95˚C. For strand hybridization, the melting temperature is 55˚C. What do you predict about the average duration of H bonds at the high temperature in comparison to the low temperature?arrow_forwardIf it is not possible to have three bands for one STR region when looking at the DNA from one individual, then how many different STR regions were sampled in the gel electrophoresis simulation to end up with the three bands on the gel. Note: there are several possible correct answers to this question.arrow_forwardFor each of the following ( A & B ) provide the method of transfer and a brief explanation as to why the method would not take place under the conditions described . 1. Which method of DNA transfer between bacteria would not take place if the donor and recipient were separated by a filter with a pore size of 0.45 um or another physical barrier 2. Which method of transfer would be blocked by the presence of high concentrations of DNAase ( enzymes capable of degrading DNA ) ?arrow_forward
- What will be the Tm of DNA that has cytosine plus guanine content of 30% compared to that of DNA that has a cytosine plus guanine content of 50% if both are heated under the same experimental conditions? lower higher same impossible to determine from information givenarrow_forwardBoth 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.arrow_forwardBelow is an EMSA showing four different reactions, A-D. In each tube there is some combination of labelled DNA probe, Protein X (the protein you are studying), and an antibody for Protein X. Identify which combination of components are found in each of the four reactions and explain how you determined that based on the molecular interactions being studied and your knowledge of gel electrophoresis. It is possible that multiple lanes have the same component(s). A B C D EMSAarrow_forward
- The E. coli genome has a superhelical density in vivo of about 0.06. Assuming the DNA has 10.5 bp/turn, what is the expected writhing number of the E. coli genome?arrow_forwardShow the calculations for the preparation of 0.01%, 0.001%, 0.0001%, 0.00001% standard DNA solutions via serial dilution from a 0.1% stock solution. Prepare 10.0 mL of 0.10% standard DNA solution using acetate buffer pH 4.6 as solvent. Prepare four different concentrations of the 0.10% standard DNA solution from step 1 via serial dilution to obtain 0.01%, 0.001%, 0.0001%, 0.00001% standard DNA solutions. Obtain seven clean and dry test tubes. Place 1.50 mL solutions in each tube as follows: Table 2.1. Set-up for the diphenylamine assay. Test tube # Content 1 (blank) acetate buffer pH 4.6 2 0.10% standard DNA 3 0.01% standard DNA 4 0.001% standard DNA 5 0.0001% standard DNA 6 0.00001% standard DNA 7 DNA extract from Part I Add 3.50 mL diphenylamine reagent to each tube, swirl each tube to thoroughly mix the contents and heat for 10 mins in a boiling water bath. Cool immediately under tap water. Read absorbances at 595 nm.…arrow_forwardThe DNA chromosome in E. coli contains approximately 4 million base pairs. The average gene contains about 1500 base pairs. Use this information to calculate the following (show all work ): a) The length in meters of this chromosome. b) The approximate number of genes in the chromosome (assuming no wasted DNA).arrow_forward
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