21 Questions
What is a characteristic of Single-Molecular Real-Time Sequencing (SMRT) that allows for accurate sequencing?
Parallelized sequencing and fluorescence detection
Which of the following is a limitation of Nanopore Sequencing?
Difficulty in extracting and delivering high-molecular weight DNA
What is an application of Single-Molecular Real-Time Sequencing (SMRT) in medicine?
Detecting drug resistance in Chronic myeloid leukemia
What was 454 sequencing used for, despite its high error rates and high cost?
Sequencing microbial genomes and investigating the decline of honey bee populations
What is an advantage of Nanopore Sequencing over other sequencing technologies?
Lower cost compared to other sequencing technologies
What is a limitation of Polony sequencing?
It generates a lot of raw data
What is a characteristic of Third-Generation Sequencing technologies?
Single molecule sequencing
What is a characteristic of Illumina technology?
It is highly accurate and cost-effective
What is an application of Nanopore Sequencing in agriculture?
Understanding gene structures in rice species
What is NOT a use of 454 sequencing?
Sanger sequencing
What is a strength of Polony sequencing?
It is cost-effective and accurate
What is a limitation of Single-Molecular Real-Time Sequencing (SMRT)?
Polymerase longevity affecting read length
Why is Illumina technology popular among scientists?
It is highly accurate and cost-effective
What is a feature of Nanopore Sequencing that allows for direct detection of nucleotides?
Measuring electrical current changes as DNA molecules pass through a nanopore
What is the primary advantage of High-throughput Sequencing (HTS) over traditional Sanger sequencing?
Significantly reduced cost and time required for sequencing
Which technique is commonly used in Massively Parallel Sequencing (MPSS) to label genetic material?
Fluorescent linker
What is another name for Massively Parallel Sequencing (MPSS)?
Next-generation sequencing
What is a key application of Massively Parallel Sequencing (MPSS)?
Screening entire genomes for new mutations or pathogens that may cause human disease
What is the main difference between 454 sequencing and traditional Sanger sequencing?
454 sequencing uses a special enzyme to cut genetic material, while Sanger sequencing uses PCR
Which of the following is NOT an application of Massively Parallel Sequencing (MPSS)?
Studying the genetic code of extinct species
What is the primary goal of High-throughput Sequencing (HTS) in terms of disease diagnosis?
Diagnosing diseases more quickly and accurately
Study Notes
Third-Generation Sequencing
- Allows for much longer reads (over 10,000 base pairs), single molecule sequencing, de novo assemblies of many microbial genomes, and reconstructions of animal and plant genomes
Single-Molecular Real-Time Sequencing (Pacific Biosciences)
- Sequences DNA in real-time, uses long DNA molecules, and combines parallelized single molecule sequencing and fluorescence detection
- Applications: environment research, medicine, and agriculture
- Advantages: high sequencing accuracy, long read lengths (up to 30 kb), and enables full gene isoforms and splice variants to be sequenced
- Limitations: polymerase longevity affects read length
Nanopore Sequencing (Oxford Nanopore Technologies)
- Uses a device called Oxford Nanopore MinION, measures electrical current changes as DNA molecules pass through a nanopore, and directly detects nucleotides without DNA synthesis
- Advantages: long read lengths (up to 2.3 Mb), lower cost (around $1000), and enables de novo genome assembly and determination of genome structural variations
- Limitations: difficulty in extracting and delivering high-molecular weight DNA, higher error rate and lower accuracy compared to short-read sequencing, and requires correction using short-read sequencing data
- Useful for: large complex genomes, identifying pathogens in clinical samples, and comparing microbial composition in soil samples
High-Throughput Sequencing (HTS)
- Based on pyrosequencing and PCR (polymerase chain reaction)
- Revolutionized the field of genetics, making it possible to sequence entire genomes in weeks instead of years and reducing the cost of sequencing from billions of dollars to just $600
- Transformed many fields, including science, medicine, and research
Massively Parallel Sequencing (MPSS)
- A powerful technology that allows for high-throughput genetic sequencing at a lower cost
- Also known as next-generation sequencing and has revolutionized the field of genomics
- Many applications, including: discovering new genes in plants, screening entire genomes for new mutations or pathogens that may cause human disease
454 Sequencing
- First next-generation sequencing (NGS) technology, developed in 2005
- Important for sequencing microbial genomes, studying metagenomics, and investigating the decline of honey bee populations
- Limitations: high error rates in certain regions, higher cost compared to other technologies, and discontinued in 2016
Polony Sequencing
- A genetic sequencing technique developed in the 1990s and 2000s, cost-effective and accurate
- Uses a special film to amplify genetic material, making it possible to: detect rare genetic messages, identify changes in gene expression, and sequence genomes of bacteria
- While it generates a lot of raw data, it's a low-cost and highly accurate technique
Illumina Technology
- A powerful tool for genetic sequencing, dominating the field since 2007
- Highly accurate and cost-effective, making it a popular choice for scientists
This quiz covers the development of third-generation sequencing, specifically Pacific Biosciences' Single-Molecule Real-Time Sequencing technology, allowing for longer reads and de novo assemblies of genomes.
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