Microbiology: An Introduction
Microbiology: An Introduction
12th Edition
ISBN: 9780321929150
Author: Gerard J. Tortora, Berdell R. Funke, Christine L. Case
Publisher: PEARSON
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Chapter 9, Problem 1CAE
Summary Introduction

Introduction:

Polymerase chain reaction (PCR) is an in vitro amplification technique implemented to generate many copies of a given DNA fragment.

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Pipetting 1. Explain why a micropipettor is an important instrument in biochemistry labs. 2. Describe the basic components (and function) of a micropipettor. Bacterial Techniques 1. Define the following. (a) Serial Dilution (b) Streak Plating (c) Spread Plating Transformation 1. Define bacterial transformation and explain why it is an important method in biochemistry labs. 2. Describe (figure, narrative) a plasmid and describe the basic components of a plasmid. 3. What role does CaCl2 play in bacterial transformation? 4. What role does heat shock play in bacterial transformation? Plasmid Isolation 1. Describe the roles the following play in plasmid purification. (a) Lysis Buffer (b) Neutralization Buffer (c) Elution Buffer
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2. Restriction Enzyme Mapping - use any resources to assist you including the hints below. A circular plasmid molecule (12,000 total base pairs in size) was cut with a series of restriction enzymes and the digestions were size fractionated by agarose gel electrophoresis. Some digestions involved just one enzyme (single digest), some combinations of two enzymes (double digests), and one utilized all three enzymes (triple digest). Agarose gel electrophoresis of the digestions produced bands of the following sizes. Enzyme EcoRI Hindi!! Pstl EcoRI and Hindill EcoRI and Psti Hindill and Pstl EcoRI, Hindill, and Pst I Bands of the Agarose Gel (size in base pairs) 2300 and 9700 4000 and 8000 12,000 800, 1500, 2500, and 7200 2300, 3200, and 6500 4000 800, 1500, 2500, 3200, and 4000 I Draw a plasmid map showing the location of the restriction enzyme sites relative to each other for each map. Include all 7 maps in your answer.
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