Genetic Analysis: An Integrated Approach (2nd Edition)
Genetic Analysis: An Integrated Approach (2nd Edition)
2nd Edition
ISBN: 9780321948908
Author: Mark F. Sanders, John L. Bowman
Publisher: PEARSON
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Chapter 10, Problem 12P
Summary Introduction

To analyze:

Why the positions of nucleic acid bands observed in one kind of blot cannot be directly compared with those in the other.

Introduction:

Blotting is the technique routinely used for the detection of biomolecules such as DNA, RNA, and proteins, named as Southern blotting, northern blotting, and western blotting respectively. In southern and northern blot method, the DNA and RNA are separated by agarose gel electrophoresis and then detected by using the DNA probe complementary to the targeted sample. In autoradiography, the radioactively labelled substances with radioisotopes are used. In a bio-analytical procedure, such substances that are distributed in a biological sample are visualized.

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Describe the steps, starting from an endonuclease digested DNA sample, to complete a Southern blot experiment.  What difference would a Northern blot have?
A Western blot is seen in the picture above. Sodium dodecyl sulfate gel polyacrylamide gel electrophoresis was used to isolate the proteins (SDS-PAGE).1. Since gel electrophoresis has isolated individual proteins, what kind of molecule is used to detect them? 2. On the right side of this Western blot, three molecules are identified. One of the bands corresponds to the smallest molecule?
Describe what each of the 12 bands on your Western blot should have been. Remember, the 12 bands will be 4 conditions x 3 proteins (phospho-S6, phospho-AMPK, tubulin). Please describe the relative density of each band compared to the control (for example, how dense will phospho-S6 be in each of the three experimental conditions compared to the control condition?). For each band, provide a well-reasoned rationale for your anticipated result. Give the reason why cell signaling would produce each result in each condition.
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