18. For DNA sequencing using the Sanger Method and radioactivity, four separate reactions (A, G, Cand T) are performed and the products of the reaction are run out on a polyacrylamide gel.For the "T" reaction:(a) List all the components that are added to the test tube.(b) Identify the component that is radioactively labeled.

Introduction: The DNA sequencing method was developed by Frederick Sanger and his colleagues in the…

Answered: 18. For DNA sequencing using the Sanger…

Biochemistry

9th Edition

ISBN:9781319114671

Author:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.

Publisher:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.

Chapter1: Biochemistry: An Evolving Science

Section: Chapter Questions

Problem 1P

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40) What external factor (aka, damaging agent) gives rise to the following DNA mutations? Write and DRAW the structure of these external damaging agents. A) interstrand crosslinks CH2 HN NH H2N NH2 DNA DNA B) 3-methyladenine 3-meA NH2 CH3 C) 1, N2-etheno-dG N- dR 1,N2-E-dGuo
a.) Explain briefly how silica allows us to purify DNA. Be sure to state the relevant biochemical features of silica and DNA as well as any relevant reagents or steps that are important for the purification. Your answer should explain both how we get the DNA to bind in the first place, and how we are able to remove the DNA. You do not (and should not) explain earlier steps in a DNA purification; focus you answer on the silica:DNA interaction. b.) In theory, can a silica spin column also be used to purify RNA? If so, explain how (and why we did not observe RNA when analyzing plasmid DNA via agarose gel electrophoresis). If not, explain why not.
After properly troubleshooting, you finally are able to visualize the gel. You obtained the image below. The ladder used is “Lambda Hind III”.Knowing that the ladder “Lambda Hind III” consists of six fragments (23130 bp, 9416 bp, 6557 bp, 4361 bp, 2322 bp, 2027 bp), properly label the bands in the gel image.
The Evidence shows which suspect was at the crime scene? ( image of murder investigation) A) suspect A B) suspect B C) suspect C D) none were at the crime scene Which of the following choices most likely lists the sizes of the four DNA fragments in lane 6 ( second image) of this agarose gel stained with ethidium bromide? ( Note: the well that the DNA was loaded into is at the top of the gel image). A) 100 bp, 300 bp, 450 bp, 600 bp. B) 300 bp, 400 bp, 100 bp, 600 bp C) 600 bp, 450 bp, 300 bp, 100 bp D) 400 bp, 300 bp, 600 bp, 100 bp E) DNA fragment size can't be estimated using an agarose gel.
H2
What volume of 4X loading buffer must be added to 21 micro L of DNA in a technique for DNA sample preparation for agarose gel electrophoresis to generate a 1X buffer solution?
1) Briefly outline the steps of DNA extraction as carried out in a lab for the purposes of sequencing, with reference to a published protocol from a company that supplies regents and kits for DNA extraction (e.g Qiagen, ThermoFischer, Invitrogen, or similar). Please Include the weblink for the published protocol you used as a reference. (Word limits: 300) 2) Why was there a greater A260 absorbance reading for your DNA sample that was incubated at higher temperatures. (Word Limits: 100)
Dr. Wakefield would like to isolate recombinant plasmids from her bacterial culture using the alkaline lysis method. She is planning to use the chemicals as listed below: Solution I: 50 mM glucose, 25 mM Tris-Cl (pH 8.0), 10 mM EDTA (pH 8.0) Solution II: ??? Solution III: 5 M potassium acetate, glacial acetic acid, de-ion water Ethanol 70% (v/v) Isopropanol TE-RNAase pH 8.0 (i) (ii) (iii) Based on the chemical list above, state the content(s) of Solution II. Explain the functions of Solution II described in Q3 a) (i) in plasmid isolation. What is the role of alcohol precipitation conducted after the plasmids are obtained at the end of the procedure? Discuss the roles of ethanol and salt in alcohol precipitation step.
123.Explain how ethidium bromide binds to the DNA and how this chemical makes DNA visible?
You are trying to determine the base content for a number of samples in the lab (mouse DNA, bacterial DNA, single-stranded DNA virus and double-stranded RNA virus). Unfortunately, you failed to use an alcohol-proof marker and during the ethanol precipitation of your samples the labels came off the tubes. You finish the experiment anyway and try to use the data to determine the samples. Can you determine the source of each sample based on your results? Briefly explain how you come to your designations. Sample % adenine % cytosine % guanine % thymine % uracil A 18 32 32 18 0 B 26 18 32 24 0 C 31 19 19 0 31 D 10 40 40 10 0
Identify the false statements. There may be more than one correct answer. A) The binding of DNA to a spin column membrane is altered by elution buffer. B) High quality DNA must be pure. C) DNA that is broken into very small pieces can still be high quality. D) Pure DNA must also be of high quality.
You have purified plasmid DNA, and observed the following absorbances: A260 = 2.9 A280 = 2.2 If A260 of 1.0 = 50 µg/ml pure dsDNA. Which of the following can you conclude? Select all that apply The concentration of your plasmid DNA is 145 ng/ul. The concentration of your plasmid DNA is 110 ng/ul. The plasmid DNA purity is acceptable. The plasmid DNA purity is not acceptable.

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