Podcast
Questions and Answers
What is the ultimate destination for non-coding RNAs?
What is the ultimate destination for non-coding RNAs?
All eukaryotic genes are encoded by RNA.
All eukaryotic genes are encoded by RNA.
False
Name two examples of non-coding RNAs.
Name two examples of non-coding RNAs.
tRNAs and rRNAs
Human genes transcribed into non-coding RNAs may equal or even exceed the number of __________ genes.
Human genes transcribed into non-coding RNAs may equal or even exceed the number of __________ genes.
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Match the following types of RNA with their characteristics:
Match the following types of RNA with their characteristics:
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Which of the following statements is true regarding non-coding RNAs?
Which of the following statements is true regarding non-coding RNAs?
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The term 'non-coding RNAs' can be shortened to 'ncRNAs'.
The term 'non-coding RNAs' can be shortened to 'ncRNAs'.
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What percentage of human genes are estimated to be non-protein-coding?
What percentage of human genes are estimated to be non-protein-coding?
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How much of the genome is considered active?
How much of the genome is considered active?
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Noncoding RNAs are translated into proteins.
Noncoding RNAs are translated into proteins.
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What is the transcriptome?
What is the transcriptome?
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The ______ project has shown that most of the non-coding DNA has a purpose.
The ______ project has shown that most of the non-coding DNA has a purpose.
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Which of the following statements about noncoding RNAs is false?
Which of the following statements about noncoding RNAs is false?
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Match the following types of non-coding RNAs with their descriptions:
Match the following types of non-coding RNAs with their descriptions:
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What does qPCR stand for?
What does qPCR stand for?
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Microarrays can measure gene expression for thousands of genes at a time.
Microarrays can measure gene expression for thousands of genes at a time.
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What is transcriptomics?
What is transcriptomics?
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High throughput RNA ________ allows for sequencing of all transcripts without limits.
High throughput RNA ________ allows for sequencing of all transcripts without limits.
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Which of the following is a limitation of qPCR?
Which of the following is a limitation of qPCR?
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Match the following techniques with their key features:
Match the following techniques with their key features:
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Transcriptomics is limited to a known number of genes.
Transcriptomics is limited to a known number of genes.
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Microarrays focus primarily on _______ coding genes.
Microarrays focus primarily on _______ coding genes.
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What is the purpose of using fluorescently labelled dNTPs in sequencing by synthesis?
What is the purpose of using fluorescently labelled dNTPs in sequencing by synthesis?
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RNA-seq is less effective at quantifying highly expressed genes compared to microarrays.
RNA-seq is less effective at quantifying highly expressed genes compared to microarrays.
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Explain one advantage of RNA-seq over microarrays.
Explain one advantage of RNA-seq over microarrays.
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A primer anneals to the linker at the end of the cDNA fragment attached to the __________.
A primer anneals to the linker at the end of the cDNA fragment attached to the __________.
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Match the following RNA-seq advantages with their descriptions:
Match the following RNA-seq advantages with their descriptions:
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What happens when the correct dNTP is incorporated during sequencing by synthesis?
What happens when the correct dNTP is incorporated during sequencing by synthesis?
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The chain-terminating modification of dNTPs in sequencing by synthesis cannot be reversed.
The chain-terminating modification of dNTPs in sequencing by synthesis cannot be reversed.
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What is a challenge of working with RNA samples?
What is a challenge of working with RNA samples?
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What is the main purpose of RNA-sequencing?
What is the main purpose of RNA-sequencing?
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RNA-seq can provide sequence level information that microarrays cannot.
RNA-seq can provide sequence level information that microarrays cannot.
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What format are RNA-sequencing outputs typically in?
What format are RNA-sequencing outputs typically in?
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RNA-sequencing can yield up to ______ million reads.
RNA-sequencing can yield up to ______ million reads.
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Match the following terms with their descriptions:
Match the following terms with their descriptions:
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Which of the following is an advantage of RNA-seq over microarrays?
Which of the following is an advantage of RNA-seq over microarrays?
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Short sequence reads from RNA-seq are typically 150-200 bp long.
Short sequence reads from RNA-seq are typically 150-200 bp long.
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What is the minimum length of short sequence reads in RNA-seq?
What is the minimum length of short sequence reads in RNA-seq?
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Study Notes
Non-Coding RNAs
- Human genes transcribed into non-protein-coding RNAs may equal or exceed protein-coding genes.
- Non-coding RNAs include tRNAs, rRNAs, miRNAs, and lncRNAs.
- All eukaryotic genes are encoded by DNA; non-coding RNAs are not translated into proteins.
- Previously considered "junk DNA," non-coding DNA has known functions as demonstrated by the ENCODE project.
Transcriptomics
- Transcriptomics studies gene expression by quantifying all RNAs in a specific cell or tissue at a given time.
- Unlike earlier methods, RNA-seq provides a comprehensive analysis without prior gene knowledge.
High Throughput Sequencing
- RNA-seq uses high throughput sequencing to analyze RNA from cells or tissues.
- Sequencing by synthesis involves primer annealing, polymerase extension, and fluorescent labeling of dNTPs.
Advantages of RNA-seq
- RNA-seq identifies approximately 40% more transcripts than microarrays and quantifies low and high expressed genes effectively.
- Can detect alternative splicing, novel exons, and mutations in transcripts.
- Provides a highly sensitive and dynamic range of data.
Challenges in RNA Transcriptome Studies
- RNA samples are fragile and can be unstable, impacting data accuracy and reliability.
- Highly expressed RNAs may dominate sequencing resources, limiting analysis of less abundant transcripts.
RNA-sequencing Process
- The RNA-seq process includes three main steps: library preparation, clonal amplification, and sequencing.
- A fastq format is utilized for storing sequencing data, containing millions of short sequence reads.
Key Takeaways from Part 2 Summary
- RNA-sequencing offers an unbiased quantitative survey of transcripts, including novel RNA detection.
- Compared to microarrays, RNA-seq provides more detailed sequence-level information.
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Description
Explore the fascinating world of non-coding RNAs and transcriptomics in this quiz. Learn about the roles of different non-coding RNAs, the advancements in RNA-seq technology, and the advantages it holds over traditional methods. Test your understanding of how these elements contribute to gene expression studies.
