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RNA Sequencing Process and Applications-F19960606001

RNA sequencing

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Ravindu Dissanayake
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35 views7 pages

RNA Sequencing Process and Applications-F19960606001

RNA sequencing

Uploaded by

Ravindu Dissanayake
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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RNA SEQUENCING PROCESS AND F19960606001

APPLICATIONS Indragi Ekanayake


INTRODUCTION TO RNA SEQUENCING
(RNA-SEQ)
Definition: RNA sequencing (RNA-Seq) is a powerful technology used to study the
transcriptome, the complete set of RNA transcripts produced by the genome.
Overview:Developed as a high-throughput sequencing method.Allows for the precise
measurement of gene expression.Facilitates the discovery of novel transcripts, splice
variants, and post-transcriptional modifications.
Key Benefits:Unbiased approach.High sensitivity and accuracy.
RNA-SEQ PROCESS OVERVIEW
1. Sample Preparation:Isolate total RNA or mRNA from cells or tissues.Remove ribosomal RNA
(rRNA) for enriched mRNA samples.
2. cDNA Synthesis:Convert RNA into complementary DNA (cDNA) using reverse transcription.
3. Library Construction:Prepare cDNA library with adapters for sequencing.
4. Sequencing:Use high-throughput sequencing (e.g., Illumina, PacBio) to read sequences.
5. Data Analysis:Align RNA-Seq reads to a reference genome or transcriptome.Quantify
expression levels, identify gene variants, and detect novel transcripts.
APPLICATIONS OF RNA SEQUENCING
❖Gene Expression Profiling:Quantify expression levels across different tissues or
conditions (e.g., disease vs. healthy).
❖Disease Mechanism Understanding:Study gene expression changes in diseases like
cancer, diabetes, or neurodegenerative disorders.
❖Single-cell RNA-Seq (scRNA-Seq):Examine gene expression at the single-cell level to
understand cellular heterogeneity.
❖RNA Editing and Splicing:Discover novel splicing variants and RNA editing sites.
❖Drug Development:Use RNA-Seq data for identifying therapeutic targets and drug
responses.
ADVANTAGES AND CHALLENGES OF RNA-SEQ
Advantages:
Unbiased Approach: Captures all RNA species without prior knowledge.High
Sensitivity: Detects rare transcripts and low-abundance RNA.Wide Applications:
Versatile for various fields like oncology, neurology, and immunology.
Challenges
Complex Data Analysis: Requires advanced bioinformatics tools and
expertise.Cost: High cost of sequencing platforms and computational
resources.Sample Quality: RNA is highly susceptible to degradation, requiring
careful handling.
CONCLUSION AND FUTURE PERSPECTIVES
Summary:RNA-Seq is a versatile tool for transcriptomics research. Revolutionized our
understanding of gene regulation, splicing, and disease mechanisms.
Future Trends:Increasing use in precision medicine. Advancements in scRNA-Seq and
long-read RNA sequencing technologies.
Closing Thoughts:RNA-Seq will continue to play a crucial role in biology, medicine,
and drug discovery.
REFERENCES
Wang, Z., Gerstein, M., & Snyder, M. (2009). RNA-Seq: a revolutionary tool for
transcriptomics. Nature Reviews Genetics, 10(1), 57-63.
Stark, R., Grzelak, M., & Hadfield, J. (2019). RNA sequencing: the teenage years.
Nature Reviews Genetics, 20(11), 631-656.
Ozsolak, F., & Milos, P. M. (2011). RNA sequencing: advances, challenges, and
opportunities. Nature Reviews Genetics, 12(2), 87-98.
Tang, F., et al. (2009). mRNA-Seq whole-transcriptome analysis of a single cell.
Nature Methods, 6(5), 377-382.
Conesa, A., et al. (2016). A survey of best practices for RNA-Seq data analysis.
Genome Biology, 17(1), 13.

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