COPY: DNA Sequencing: Sanger Method Overview

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What is the primary purpose of DNA sequencing?

To determine the order of bases in DNA (correct)

To identify different types of RNA

To isolate DNA from bacterial cells

To amplify DNA through PCR

Which of the following best describes the Sanger sequencing method?

It uses multiple primers for DNA synthesis.

It requires a complex array of enzymes to function.

It is classified as a first generation sequencing method. (correct)

It is a second generation sequencing technique.

What are ddNTPs in the context of Sanger sequencing?

Nucleotides that lack phosphate groups

Standard nucleotides used in PCR amplification

Nucleotides that promote continuous DNA synthesis

Fluorescently labeled nucleotides that cause chain termination (correct)

What role does the single primer play in the Sanger sequencing process?

It initiates DNA synthesis from the 3’ end of the template. Signup and view all the answers

Which advancement in genetic research has DNA sequencing contributed to significantly?

The identification of mutations associated with diseases Signup and view all the answers

What is the first step of the Sanger sequencing method?

DNA denaturation and template preparation Signup and view all the answers

Who developed the Sanger sequencing method?

Frederick Sanger and his colleagues Signup and view all the answers

What is a significant difference between dNTPs and ddNTPs?

ddNTPs lack the 3’ hydroxyl group required for DNA extension. Signup and view all the answers

What is the role of labelled dideoxynucleotides (ddNTPs) in the Sanger sequencing method?

To terminate the growing DNA chain Signup and view all the answers

Which of the following statements describes an outcome of capillary electrophoresis in Sanger sequencing?

Each amplicon is separated based on its length. Signup and view all the answers

What components are included in the mixture for preparing template DNA in Sanger sequencing?

A mixture of dNTPs and ddNTPs Signup and view all the answers

Why are smaller amounts of ddNTPs used compared to dNTPs in the sequencing process?

To ensure that chain termination occurs at various lengths Signup and view all the answers

What is the primary function of the fluorescent tag on the dideoxynucleotides in Sanger sequencing?

To generate a chromatogram Signup and view all the answers

During the Sanger sequencing process, what directly excites the labels on the nucleotides?

A laser Signup and view all the answers

What is the result seen in the chromatogram generated by Sanger sequencing?

A sequence of peaks corresponding to different nucleotides Signup and view all the answers

In Sanger sequencing, what happens once a dideoxynucleotide is incorporated into the growing DNA chain?

The chain is terminated and no further bases can be added Signup and view all the answers

What characterized the Human Genome Project?

It was completed in 2003 using Sanger sequencing. Signup and view all the answers

Next Generation Sequencing was first introduced in 2000.

False Signup and view all the answers

What is one major benefit of Next Generation Sequencing compared to first-generation methods?

Increased speed and accuracy Signup and view all the answers

Next Generation Sequencing is sometimes referred to as __________ sequencing.

second-generation Signup and view all the answers

Match the following NGS categories with their descriptions:

Sequencing by hybridization = Uses specific probes to interrogate sequences Sequencing by Synthesis = Further development of Sanger sequencing RNA sequencing (RNA-seq) = Quantify mRNAs for gene expression analysis Deep sequencing = Rapidly sequence whole genomes Signup and view all the answers

Which of the following is an application of NGS?

Analysing epigenetic factors Signup and view all the answers

Next Generation Sequencing allows the identification of novel RNA variants.

True Signup and view all the answers

What is the purpose of adapter sequences in library preparation?

To provide a universal priming site for sequencing primers Signup and view all the answers

Sequencing machines amplify each library fragment on a solid surface, such as beads or flat silicon.

True Signup and view all the answers

What is a common characteristic of Sequencing by Synthesis (SBS)?

Relies on shorter reads and has higher error rates compared to Sanger sequencing. Signup and view all the answers

The raw data provided by sequencing machines consists of a collection of _____________ generated at each cluster.

DNA sequences Signup and view all the answers

Match the following properties with their relevance in sequencing:

Shorter reads (300-500 bp) = Sequencing by Synthesis (SBS) Higher error rate = Sequencing by Synthesis (SBS) Massively parallel sequencing = High sequence coverage Reversibly fluorescent nucleotides = Step-by-step nucleotide incorporation Signup and view all the answers

What happens to each cluster during the sequencing process?

Each cluster acts as an individual sequencing reaction Signup and view all the answers

Sequencing by Synthesis uses only one type of nucleotide for DNA sequencing.

False Signup and view all the answers

What is the significance of high sequence coverage in Sequencing by Synthesis?

It allows for the generation of millions to billions of short DNA sequence reads. Signup and view all the answers

Sequencing by Synthesis relies on _____________ incorporation of nucleotides.

step-by-step Signup and view all the answers

What is the primary function of reversible termination in nucleotide sequencing?

To ensure only one nucleotide is incorporated per cycle Signup and view all the answers

In Illumina sequencing, PCR amplification is performed after the library preparation step.

True Signup and view all the answers

What is the typical length of DNA fragments after fragmentation during library preparation?

200 – 600 bp Signup and view all the answers

The process of ________ occurs to bind complementary DNA to primers on the surface of the flow cell.

washing Signup and view all the answers

Match the following NGS technologies with their characteristics:

454 sequencing = Utilizes pyrosequencing Illumina = Cyclic reversible termination SOLiD = Uses ligation-based sequencing Ion Torrent = Measures pH changes during sequencing Signup and view all the answers

During bridge amplification in Illumina sequencing, what is the main purpose of the flexible linker?

To facilitate contact between the DNA template and primers Signup and view all the answers

In the sequencing process, fluorescent signals are read only after the clustering of DNA fragments.

True Signup and view all the answers

What type of molecules are cleaved and washed away after the fluorescent signal is read?

Fluorescent molecules and terminator groups Signup and view all the answers

The first step in the Illumina sequencing process is ________ preparation.

library Signup and view all the answers

COPY: DNA Sequencing: Sanger Method Overview

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Questions and Answers

What is the primary purpose of DNA sequencing?

Which of the following best describes the Sanger sequencing method?

What are ddNTPs in the context of Sanger sequencing?

What role does the single primer play in the Sanger sequencing process?

Which advancement in genetic research has DNA sequencing contributed to significantly?

What is the first step of the Sanger sequencing method?

Who developed the Sanger sequencing method?

What is a significant difference between dNTPs and ddNTPs?

What is the role of labelled dideoxynucleotides (ddNTPs) in the Sanger sequencing method?

Which of the following statements describes an outcome of capillary electrophoresis in Sanger sequencing?

What components are included in the mixture for preparing template DNA in Sanger sequencing?

Why are smaller amounts of ddNTPs used compared to dNTPs in the sequencing process?

What is the primary function of the fluorescent tag on the dideoxynucleotides in Sanger sequencing?

During the Sanger sequencing process, what directly excites the labels on the nucleotides?

What is the result seen in the chromatogram generated by Sanger sequencing?

In Sanger sequencing, what happens once a dideoxynucleotide is incorporated into the growing DNA chain?

What characterized the Human Genome Project?

Next Generation Sequencing was first introduced in 2000.

What is one major benefit of Next Generation Sequencing compared to first-generation methods?

Next Generation Sequencing is sometimes referred to as __________ sequencing.

Match the following NGS categories with their descriptions:

Which of the following is an application of NGS?

Next Generation Sequencing allows the identification of novel RNA variants.

What is the purpose of adapter sequences in library preparation?

Sequencing machines amplify each library fragment on a solid surface, such as beads or flat silicon.

What is a common characteristic of Sequencing by Synthesis (SBS)?

The raw data provided by sequencing machines consists of a collection of _____________ generated at each cluster.

Match the following properties with their relevance in sequencing:

What happens to each cluster during the sequencing process?

Sequencing by Synthesis uses only one type of nucleotide for DNA sequencing.

What is the significance of high sequence coverage in Sequencing by Synthesis?

Sequencing by Synthesis relies on _____________ incorporation of nucleotides.

What is the primary function of reversible termination in nucleotide sequencing?

In Illumina sequencing, PCR amplification is performed after the library preparation step.

What is the typical length of DNA fragments after fragmentation during library preparation?

The process of ________ occurs to bind complementary DNA to primers on the surface of the flow cell.

Match the following NGS technologies with their characteristics:

During bridge amplification in Illumina sequencing, what is the main purpose of the flexible linker?

In the sequencing process, fluorescent signals are read only after the clustering of DNA fragments.

What type of molecules are cleaved and washed away after the fluorescent signal is read?

The first step in the Illumina sequencing process is ________ preparation.

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