Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

RNA-seq03:21

RNA-seq

9.8K
RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases. 
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while...
9.8K
Next-generation Sequencing03:00

Next-generation Sequencing

87.6K
The first human genome sequencing project cost $2.7 billion and was declared complete in 2003, after 15 years of international cooperation and collaboration between several research teams and funding agencies. Today, with the advent of next-generation sequencing technologies, the cost and time of sequencing a human genome have dropped over 100 fold.
Next-Generation Sequencing Methods
Although all next-generation methods use different technologies, they all share a set of standard features....
87.6K
Sanger Sequencing01:57

Sanger Sequencing

753.0K
DNA sequencing is a fundamental technique that is routinely used in the biological sciences. This method can be applied to a range of questions at different scales - from the sequencing of a cloned DNA fragment or the study of a mutation in a gene up to whole-genome sequencing. However, despite the widespread use of sequencing today, it was not until 1977 that Fredrick Sanger and his collaborators developed the chain-termination method to decode DNA sequences. It relies on the separation of a...
753.0K

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Inflammasome Activation Differences Underpin Different <i>Mycobacterium tuberculosis</i> Infection Outcomes.

MedComm·2025
Same author

Inflammasome activation differences underpin different <i>Mycobacterium tuberculosis</i> infection outcomes.

bioRxiv : the preprint server for biology·2025
Same author

Initial Analysis of Plant Soil for Evidence of Pathogens Associated with a Disease of Seedling <i>Ocotea monteverdensis</i>.

Microorganisms·2025
Same author

Intranasal Leukemia Inhibitory Factor as a late-stage treatment for delayed white matter damage in concussive head injury.

bioRxiv : the preprint server for biology·2025
Same author

Correction to: Traumatic brain injury-induced downregulation of Nrf2 activates inflammatory response and apoptotic cell death.

Journal of molecular medicine (Berlin, Germany)·2025
Same author

Use of a logging road in a Costa Rican forest changes the composition and stability of soil microbial decomposer communities, and the conversion of organic carbon into biomass.

Journal of applied microbiology·2025

Related Experiment Video

Updated: Jun 7, 2025

3' End Sequencing Library Preparation with A-seq2
12:01

3' End Sequencing Library Preparation with A-seq2

Published on: October 10, 2017

10.5K

RNA Sequencing Protocols for Short-Read Sequencing.

Laura Vasquez-Velez1, Veera D'Mello1, Patricia Soteropoulos2

  • 1Genomics Center, Rutgers New Jersey Medical School, Newark, NJ, USA.

Methods in Molecular Biology (Clifton, N.J.)
|November 15, 2024
PubMed
Summary

RNA sequencing (RNA-seq) enables discovery of new RNA variants. This study details four library preparation protocols for short-read sequencing, including poly(A) selection, ribosomal depletion, and SMART technology for low-input samples.

Keywords:
Library preparationPoly(A) selectionRNA-seqRibosomal depletionShort-read sequencingcDNA

More Related Videos

Rare Event Detection Using Error-corrected DNA and RNA Sequencing
10:36

Rare Event Detection Using Error-corrected DNA and RNA Sequencing

Published on: August 3, 2018

12.0K
Improving Small RNA-seq: Less Bias and Better Detection of 2'-O-Methyl RNAs
08:49

Improving Small RNA-seq: Less Bias and Better Detection of 2'-O-Methyl RNAs

Published on: September 16, 2019

7.6K

Related Experiment Videos

Last Updated: Jun 7, 2025

3' End Sequencing Library Preparation with A-seq2
12:01

3' End Sequencing Library Preparation with A-seq2

Published on: October 10, 2017

10.5K
Rare Event Detection Using Error-corrected DNA and RNA Sequencing
10:36

Rare Event Detection Using Error-corrected DNA and RNA Sequencing

Published on: August 3, 2018

12.0K
Improving Small RNA-seq: Less Bias and Better Detection of 2'-O-Methyl RNAs
08:49

Improving Small RNA-seq: Less Bias and Better Detection of 2'-O-Methyl RNAs

Published on: September 16, 2019

7.6K

Area of Science:

  • Molecular Biology
  • Genomics
  • Bioinformatics

Background:

  • RNA sequencing (RNA-seq) is crucial for identifying novel RNA variants and transcripts.
  • Standard RNA-seq involves RNA capture, complementary DNA (cDNA) conversion, and cDNA modification for sequencing.

Purpose of the Study:

  • To describe and compare four distinct library preparation protocols for short-read RNA sequencing.
  • To evaluate methods suitable for varying sample input amounts, including low and pico quantities.

Main Methods:

  • Poly(A) selection for capturing messenger RNA (mRNA).
  • Ribosomal RNA (rRNA) depletion for removing abundant rRNA.
  • SMART (Switching Mechanism at 5' end of RNA Template) technology for cDNA synthesis, particularly for low-input samples.

Main Results:

  • Successful implementation of four distinct library preparation workflows.
  • Demonstration of the utility of SMART technology for low and pico RNA inputs.
  • Comparison of protocol efficiency for different RNA species capture and modification strategies.

Conclusions:

  • The described protocols offer versatile options for RNA-seq library preparation.
  • Specific methods are optimized for different RNA types and input amounts, enhancing experimental flexibility.
  • These protocols facilitate robust discovery of RNA variants and transcripts across diverse biological samples.