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Questions and Answers
What is the role of dideoxynucleotides in Sanger sequencing?
What is the role of dideoxynucleotides in Sanger sequencing?
- To terminate DNA strand elongation. (correct)
- To provide energy for DNA polymerase.
- To stabilize the DNA template.
- To initiate DNA replication.
In Sanger sequencing, what does each test tube contain in addition to the DNA template, DNA polymerase, and DNA primers?
In Sanger sequencing, what does each test tube contain in addition to the DNA template, DNA polymerase, and DNA primers?
- All four types of deoxynucleotides (A, C, T, G).
- One type of dideoxynucleotide (A*, C*, T*, or G*) and all four types of deoxynucleotides (A, C, T, G). (correct)
- Only one type of deoxynucleotide.
- All four types of dideoxynucleotides (A*, C*, T*, G*).
What is the purpose of gel electrophoresis in Sanger sequencing?
What is the purpose of gel electrophoresis in Sanger sequencing?
- To isolate the DNA polymerase.
- To separate DNA fragments based on size. (correct)
- To amplify the DNA fragments.
- To label the DNA fragments with fluorescent markers.
What is the role of DNA polymerase in Sanger sequencing?
What is the role of DNA polymerase in Sanger sequencing?
What determines the length of the DNA fragments produced during Sanger sequencing?
What determines the length of the DNA fragments produced during Sanger sequencing?
Flashcards
High-throughput sequencing
High-throughput sequencing
Rapid sequencing methods to analyze genomes efficiently.
Dideoxynucleotide
Dideoxynucleotide
Modified nucleotides that terminate DNA replication when added.
Chain termination method
Chain termination method
Sequencing method where dideoxynucleotides stop DNA strand elongation.
Gel electrophoresis
Gel electrophoresis
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DNA polymerase
DNA polymerase
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Study Notes
High-Throughput DNA Sequencing
- High-throughput sequencing allows rapid sequencing of many genomes simultaneously.
Chain Termination Method
- Uses dideoxynucleotides (labeled with *) to stop DNA replication at specific points.
- Four separate reactions, each with a different dideoxynucleotide.
- Fragments of varying lengths are produced, depending on where the dideoxynucleotide is incorporated.
- Gel electrophoresis separates these fragments by length.
- The first base in the sequence is identified by the band furthest from the bottom of the gel, indicating the shortest fragment.
- Each subsequent base is identified in the same manner, effectively creating the sequence.
Capillary Gel Electrophoresis
- A faster version of the chain termination method.
- Dideoxynucleotides are labelled with fluorescent dyes.
- Single-stranded DNA fragments are separated by gel electrophoresis within a capillary tube.
- A laser illuminates these dyes, and a detector records the color and position.
- The data gathered is converted into a sequence.
Next-Generation Sequencing
- Newer methods avoid gel electrophoresis.
- Examples include nanopore sequencing and pyrosequencing.
- Essentially, automated systems.
Benefits of High-Throughput Sequencing
- Increased speed enables analysis of many genomes.
- Useful in fields like evolutionary biology and classification.
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Description
Explore high-throughput DNA sequencing, including the chain termination method using dideoxynucleotides to halt DNA replication. Learn about capillary gel electrophoresis, a faster iteration using fluorescent dyes and laser detection for efficient sequencing.
