Textbook Question
The following dideoxy DNA sequencing gel is produced in a laboratory.
What is the double-stranded DNA sequence of this molecule? Label the polarity of each strand.
Ch. 7 - DNA Structure and Replication
Sanders3rd EditionGenetic Analysis: An Integrated ApproachISBN: 9780135564172
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Table of contents
- Ch. 1 - The Molecular Basis of Heredity, Variation, and Evolution64
- Ch. 2 - Transmission Genetics144
- Ch. 3 - Cell Division and Chromosome Heredity81
- Ch. 4 - Gene Interaction87
- Ch. 5 - Genetic Linkage and Mapping in Eukaryotes103
- Ch. 6 - Genetic Analysis and Mapping in Bacteria and Bacteriophages73
- Ch. 7 - DNA Structure and Replication71
- Ch. 8 - Molecular Biology of Transcription and RNA Processing79
- Ch. 9 - The Molecular Biology of Translation110
- Ch. 10 - Eukaryotic Chromosome Abnormalities and Molecular Organization100
- Ch. 11 - Gene Mutation, DNA Repair, and Homologous Recombination86
- Ch. 12 - Regulation of Gene Expression in Bacteria and Bacteriophage90
- Ch. 13 - Regulation of Gene Expression in Eukaryotes38
- Ch. 14 - Analysis of Gene Function via Forward Genetics and Reverse Genetics72
- Ch. 15 - Recombinant DNA Technology and Its Applications69
- Ch. 16 - Genomics: Genetics from a Whole-Genome Perspective49
- Ch. 17 - Organelle Inheritance and the Evolution of Organelle Genomes27
- Ch. 18 - Developmental Genetics43
- Ch. 19 - Genetic Analysis of Quantitative Traits66
- Ch. 20 - Population Genetics and Evolution at the Population, Species, and Molecular Levels105
Sanders3rd EditionGenetic Analysis: An Integrated ApproachISBN: 9780135564172Not the one you use?Change textbook
Chapter 7, Problem 28b
In a dideoxy DNA sequencing experiment, four separate reactions are carried out to provide the replicated material for DNA sequencing gels. Reaction products are usually run in gel lanes labeled A, T, C, and G.
How does PCR play a role in dideoxy DNA sequencing?
Verified step by step guidance1
Understand that dideoxy DNA sequencing, also known as Sanger sequencing, relies on the selective incorporation of chain-terminating dideoxynucleotides (ddNTPs) during DNA synthesis.
Recognize that PCR (Polymerase Chain Reaction) is used to amplify the DNA template before sequencing. This ensures there is enough DNA material to work with in the sequencing reactions.
In the sequencing process, PCR is modified to include a mixture of normal deoxynucleotides (dNTPs) and a small proportion of ddNTPs, which terminate DNA synthesis when incorporated. Each ddNTP is labeled with a specific marker (e.g., a fluorescent dye) corresponding to one of the four bases (A, T, C, or G).
The PCR reactions are carried out in four separate tubes, each containing one of the four ddNTPs (ddATP, ddTTP, ddCTP, or ddGTP). This ensures that DNA fragments of varying lengths are generated, each terminating at a specific base.
The resulting DNA fragments are then separated by size using gel electrophoresis, and the sequence of the original DNA template is determined by analyzing the labeled ddNTPs in the gel lanes labeled A, T, C, and G.
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Key Concepts
Here are the essential concepts you must grasp in order to answer the question correctly.
Polymerase Chain Reaction (PCR)
PCR is a technique used to amplify specific DNA sequences, generating millions of copies from a small initial sample. This amplification is crucial for dideoxy DNA sequencing, as it ensures that there is enough DNA available for analysis. The process involves repeated cycles of denaturation, annealing, and extension, allowing for precise replication of the target DNA.
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Dideoxy Sequencing Method
Dideoxy sequencing, also known as Sanger sequencing, is a method for determining the nucleotide sequence of DNA. It involves incorporating dideoxynucleotides, which terminate DNA strand elongation, into the growing DNA strands during replication. The resulting fragments are then separated by size using gel electrophoresis, allowing for the identification of the sequence based on the labeled ends of the fragments.
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Gel Electrophoresis
Gel electrophoresis is a laboratory technique used to separate DNA fragments based on their size. In the context of dideoxy sequencing, the amplified DNA fragments produced from the sequencing reactions are loaded into a gel matrix and subjected to an electric field. Smaller fragments migrate faster than larger ones, allowing for the visualization of the different lengths of DNA strands, which correspond to the nucleotide sequence.
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Related Practice
Textbook Question
A family consisting of a mother (I-1), a father (I-2), and three children (II-1, II-2, and II-3) are genotyped by PCR for a region of an autosome containing repeats of a 10-bp sequence. The mother carries 16 repeats on one chromosome and 21 on the homologous chromosome. The father carries repeat numbers of 18 and 26.
Identify all the possible genotypes of children of this couple by specifying PCR fragment lengths in each genotype.
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Textbook Question
A family consisting of a mother (I-1), a father (I-2), and three children (II-1, II-2, and II-3) are genotyped by PCR for a region of an autosome containing repeats of a 10-bp sequence. The mother carries 16 repeats on one chromosome and 21 on the homologous chromosome. The father carries repeat numbers of 18 and 26.
What genetic term best describes the pattern of inheritance of this DNA marker? Explain your choice.
Textbook Question
In a dideoxy DNA sequencing experiment, four separate reactions are carried out to provide the replicated material for DNA sequencing gels. Reaction products are usually run in gel lanes labeled A, T, C, and G.
Identify the nucleotides used in the dideoxy DNA sequencing reaction that produces molecules for the A lane of the sequencing gel.
Textbook Question
In a dideoxy DNA sequencing experiment, four separate reactions are carried out to provide the replicated material for DNA sequencing gels. Reaction products are usually run in gel lanes labeled A, T, C, and G.
Why is incorporation of a dideoxynucleotide during DNA sequencing identified as a 'replication-terminating' event?
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Textbook Question
Using an illustration style and labeling, draw the electrophoresis gel containing dideoxy sequencing fragments for the DNA template strand 3'-AGACGATAGCAT-5'.
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