In laboratory, the following dideoxy DNA sequencing gel is produced. The double-stranded DNA sequence of this molecules to be determined. The polarity of each strand is to be labeled. Introduction: DNA sequencing is the determination of a sequence of nucleotide bases (A, T, G, C) in a DNA. Sanger sequencing is a method of sequencing a region of DNA that is up to 900 base pairs in length. This process is also known as Chain termination method. Sanger sequencing was first developed by Fred Sanger and his colleagues in 1977 . This process involved the formation of many copies of the template DNA strand. Dideoxynucleotides inhibit the chain elongation process of DNA polymerases. They are also abbreviated as 2', 3' Dideoxynucleotides (ddNTPs)- (ddATP, ddTTP, ddGTP, ddCTP). The principle of Sanger sequencing is that sufficient time and material can produce at least one DNA sequence of every possible length with a tagged nucleotide at the end. The tagged nucleotides get terminated because ddNTPs lack 3 ' − OH group that generally extends the chain.

Question

5'-[seq]-3' The diagram shows the results of gel electrophoresis for Sanger sequencing. The wells are represented by open boxes and the DNA bands are represented by black boxes. The wells are labeled to show which dideoxy reaction was loaded into each. Write the sequence of the original template strand used for this sequencing reaction, with the 5’ end on the left and the 3’ end on the right.

Answer 1

Question

Definition Definition Chemical process in which one or more monomers combine to produce a very large chain-like molecule called a polymer. The functional groups present on the monomers and their steric effects are responsible for polymerization through a sequence of reactions that vary in complexity. There exists a stable covalent chemical bond between monomers that sets apart polymerization from other processes.

Chapter 7, Problem 29P

Summary Introduction

To analyze:

In laboratory, the following dideoxy DNA sequencing gel is produced.

The double-stranded DNA sequence of this molecules to be determined.

The polarity of each strand is to be labeled.

Introduction:

DNA sequencing is the determination of a sequence of nucleotide bases (A, T, G, C) in a DNA.

Sanger sequencing is a method of sequencing a region of DNA that is up to 900 base pairs in length. This process is also known as Chain termination method.

Sanger sequencing was first developed by Fred Sanger and his colleagues in 1977.

This process involved the formation of many copies of the template DNA strand.

Dideoxynucleotides inhibit the chain elongation process of DNA polymerases. They are also abbreviated as 2', 3' Dideoxynucleotides (ddNTPs)- (ddATP, ddTTP, ddGTP, ddCTP).

The principle of Sanger sequencing is that sufficient time and material can produce at least one DNA sequence of every possible length with a tagged nucleotide at the end.

The tagged nucleotides get terminated because ddNTPs lack 3'− OH group that generally extends the chain.

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5'-[seq]-3' The diagram shows the results of gel electrophoresis for Sanger sequencing. The wells are represented by open boxes and the DNA bands are represented by black boxes. The wells are labeled to show which dideoxy reaction was loaded into each. Write the sequence of the original template strand used for this sequencing reaction, with the 5’ end on the left and the 3’ end on the right.

During agarose gel electrophoresis, why does DNA move through the gel when electric current is applied? because DNA is negatively charged because a charged chemical from the loading buffer is bound to the DNA because DNA is positively charged because DNA absorbs electricity

The short DNA shown below is to be sequenced. Using your knowledge of how the Sanger method works, in the gel diagram, draw in the bands that will appear when DNA polymerase is added to the reaction along with the four different nucleotide mixtures indicated. Note that some of these mixtures are not what would normally be used in a sequencing reaction. Dideoxynucleatides (ddNTPs) are added in relatively small amounts. The asterisk represents a radioactive label. *5' - 3'-ОН 3' – -- ACGACGCAGGACATTAGAC-5' Nucleotide mixtures: A. DATP, DTTP, dCTP, DGTP, ddTTP (given) B. DATP, ATTP, dCTP, AGTP, ddATE C. dTTP, dGTP. ACTP, ddCTP, ddATP D. DATP, dCTP, dTTP, ddGTP A в с D | || ||

Answer 2

Question

Definition Definition Chemical process in which one or more monomers combine to produce a very large chain-like molecule called a polymer. The functional groups present on the monomers and their steric effects are responsible for polymerization through a sequence of reactions that vary in complexity. There exists a stable covalent chemical bond between monomers that sets apart polymerization from other processes.

Chapter 7, Problem 29P

Summary Introduction

To analyze:

In laboratory, the following dideoxy DNA sequencing gel is produced.

The double-stranded DNA sequence of this molecules to be determined.

The polarity of each strand is to be labeled.

Introduction:

DNA sequencing is the determination of a sequence of nucleotide bases (A, T, G, C) in a DNA.

Sanger sequencing is a method of sequencing a region of DNA that is up to 900 base pairs in length. This process is also known as Chain termination method.

Sanger sequencing was first developed by Fred Sanger and his colleagues in 1977.

This process involved the formation of many copies of the template DNA strand.

Dideoxynucleotides inhibit the chain elongation process of DNA polymerases. They are also abbreviated as 2', 3' Dideoxynucleotides (ddNTPs)- (ddATP, ddTTP, ddGTP, ddCTP).

The principle of Sanger sequencing is that sufficient time and material can produce at least one DNA sequence of every possible length with a tagged nucleotide at the end.

The tagged nucleotides get terminated because ddNTPs lack 3'− OH group that generally extends the chain.

Expert Solution & Answer

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Answer 3

Question

Definition Definition Chemical process in which one or more monomers combine to produce a very large chain-like molecule called a polymer. The functional groups present on the monomers and their steric effects are responsible for polymerization through a sequence of reactions that vary in complexity. There exists a stable covalent chemical bond between monomers that sets apart polymerization from other processes.

Chapter 7, Problem 29P

Summary Introduction

To analyze:

In laboratory, the following dideoxy DNA sequencing gel is produced.

The double-stranded DNA sequence of this molecules to be determined.

The polarity of each strand is to be labeled.

Introduction:

DNA sequencing is the determination of a sequence of nucleotide bases (A, T, G, C) in a DNA.

Sanger sequencing is a method of sequencing a region of DNA that is up to 900 base pairs in length. This process is also known as Chain termination method.

Sanger sequencing was first developed by Fred Sanger and his colleagues in 1977.

This process involved the formation of many copies of the template DNA strand.

Dideoxynucleotides inhibit the chain elongation process of DNA polymerases. They are also abbreviated as 2', 3' Dideoxynucleotides (ddNTPs)- (ddATP, ddTTP, ddGTP, ddCTP).

The principle of Sanger sequencing is that sufficient time and material can produce at least one DNA sequence of every possible length with a tagged nucleotide at the end.

The tagged nucleotides get terminated because ddNTPs lack 3'− OH group that generally extends the chain.

Expert Solution & Answer

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See solution

Check out sample textbook solution

Answer 4

Question

Definition Definition Chemical process in which one or more monomers combine to produce a very large chain-like molecule called a polymer. The functional groups present on the monomers and their steric effects are responsible for polymerization through a sequence of reactions that vary in complexity. There exists a stable covalent chemical bond between monomers that sets apart polymerization from other processes.

Chapter 7, Problem 29P

Summary Introduction