Genetics: A Conceptual Approach 6E w/ SaplingPlus (Six-Month Access)
Genetics: A Conceptual Approach 6E w/ SaplingPlus (Six-Month Access)
6th Edition
ISBN: 9781319125929
Author: Benjamin A. Pierce
Publisher: W. H. Freeman
Question
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Chapter 19.2, Problem 30AQP

a.

Summary Introduction

To determine:

The piece of DNA is cut by EcoRI, the resulting fragments are separated by gel electrophoresis, and the gel is stained with ethidium bromide. Draw a picture of the bands that will appear on the gel.

Introduction:

EcoRI is type II restriction enzyme. 5'GAATTC3' is the recognition site of EcoRI and it is obtained from the bacterium Escherichia coli. It is type II enzyme that cut at specific site. Gel electrophoresis is the technique used for the separation of molecule in the gel under the influence of current. Smaller the molecule, easily does it pass through the pores. Ethidium bromide is staining dye, which stacks in the bases of nucleotide.

a.

Expert Solution
Check Mark

Explanation of Solution

Pictorial representation: The following gel shows that DNA fragments are present after the EcoRI has cleaved at 2 kb, 4 kb and 5 kb.

Genetics: A Conceptual Approach 6E w/ SaplingPlus (Six-Month Access), Chapter 19.2, Problem 30AQP , additional homework tip  1

Fig. 1: Gel electrophoresis DNA fragmentation.

After the action of enzyme EcoRI on the DNA, it will produce three fragments of length 2 kb, 4 kb and 5 kb. These bands will appear on the gel as shown in the Fig.1.

b.

Summary Introduction

To determine:

If a mutation that alters EcoRI site 1 occurs in this piece of DNA, how will the banding pattern on the gel differ from the one that drew in part a?

Introduction:

EcoRI is type II restriction enzyme. 5'GAATTC3' is the recognition site of EcoRI and it is obtained from the bacterium Escherichia coli. It is type II enzyme that cut at specific site. Gel electrophoresis is the technique used for the separation of molecule in the gel under the influence of current. Smaller the molecule, easily does it pass through the pores. Ethidium bromide is staining dye, which stacks in the bases of nucleotide.

b.

Expert Solution
Check Mark

Explanation of Solution

Pictorial representation: The following gel on that DNA fragments only two fragments of length is obtained 6 kb and 8 kb.

Genetics: A Conceptual Approach 6E w/ SaplingPlus (Six-Month Access), Chapter 19.2, Problem 30AQP , additional homework tip  2

Fig 2 Gel fragmentation

Only two fragments will obtained if the EcoRI site 1 is mutated, two fragments are 6 and 5 kb.

c.

Summary Introduction

To determine:

The difference of banding pattern on the gel that drew in part a if mutations that alter EcoRI sites 1 and 2 occur in this piece of DNA.

Introduction:

EcoRI is type II restriction enzyme. 5'GAATTC3' is the recognition site of EcoRI and it is obtained from the bacterium Escherichia coli. It is type II enzyme that cut at specific site. Gel electrophoresis is the technique used for the separation of molecule in the gel under the influence of current. Smaller the molecule, easily does it pass through the pores. Ethidium bromide is staining dye, which stacks in the bases of nucleotide.

c.

Expert Solution
Check Mark

Explanation of Solution

Pictorial representation: The gel electrophoresis after the mutation occurs at both sites.

Genetics: A Conceptual Approach 6E w/ SaplingPlus (Six-Month Access), Chapter 19.2, Problem 30AQP , additional homework tip  3

Fig. 3: Gel electrophoresis result

If mutation occur on the both sites, then there will be no cutting and if there will be no cutting then fragment will remain same.

d.

Summary Introduction

To determine:

The difference of banding pattern on the gel differ from the one that drew in part a if the 1000 bp of DNA were inserted between the two restriction sites.

Introduction:

EcoRI is type II restriction enzyme. 5'GAATTC3' is the recognition site of EcoRI and it is obtained from the bacterium Escherichia coli. It is type II enzyme that cut at specific site. Gel electrophoresis is the technique used for the separation of molecule in the gel under the influence of current. Smaller the molecule, easily does it pass through the pores. Ethidium bromide is staining dye, which stacks in the bases of nucleotide.

d.

Expert Solution
Check Mark

Explanation of Solution

Pictorial representation: The gel on that DNA fragments only two fragments of length is obtained 2 kb and 5 kb.

Genetics: A Conceptual Approach 6E w/ SaplingPlus (Six-Month Access), Chapter 19.2, Problem 30AQP , additional homework tip  4

Fig 4:  Gel fragmentation

Inserting 1000 kb of DNA between the restriction sites will increase the 4 kb fragment to 5 kb fragment, then two fragment 5 and 2 kb length will come, as shown in the fig 4.

e.

Summary Introduction

To determine:

The difference in the banding pattern on the gel differ from the one that drew in part a if 500 bp of DNA between the two restriction sites were deleted.

Introduction:

EcoRI is type II restriction enzyme. 5'GAATTC3' is the recognition site of EcoRI and it is obtained from the bacterium Escherichia coli. It is type II enzyme that cut at specific site. Gel electrophoresis is the technique used for the separation of molecule in the gel under the influence of current. Smaller the molecule, easily does it pass through the pores. Ethidium bromide is staining dye, which stacks in the bases of nucleotide.

e.

Expert Solution
Check Mark

Explanation of Solution

Pictorial representation: The gel on that DNA fragments only three fragments of length are obtained 5 kb, 3.5 kb and 2 kb.

Genetics: A Conceptual Approach 6E w/ SaplingPlus (Six-Month Access), Chapter 19.2, Problem 30AQP , additional homework tip  5

Fig 5 Gel fragmentation

Deleting 500 kb of DNA between the two restriction sites will reduce the size of the fragment, results three fragment obtained 5 kb, 3.5 kb and 2 kb, and they are shown in the fig 5.

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