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

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 microarray-based...
Ribosome Profiling02:24

Ribosome Profiling

Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique helps...

You might also read

Related Articles

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

Sort by
Same author

First draft genome and genetic diversity analysis of the economically important and vulnerable tree species Saraca asoca (Roxb.) W.J. de Wilde.

Journal of genetics·2026
Same author

Disrupting SnRK1β1A confers broad-spectrum resistance to fungal pathogens.

Trends in plant science·2026
Same author

An NLR-TF immune module under asymmetric selection shapes rice immunity and yield.

Molecular plant·2026
Same author

Dual regulation of RNase P subunit Rpp30 by an acetyltransferase and E3 ligase in rice immunity.

Plant physiology·2026
Same author

A Conserved Magnaporthe oryzae Effector Counteracts the Rice Ubiquitin-Proteasome System by Disrupting the E2 Function to Suppress Immunity.

Plant biotechnology journal·2026
Same author

Systemic Delivery of Functional Proteins Into Plants Using an Engineered Membrane Translocation Domain.

Plant biotechnology journal·2026

Related Experiment Video

Updated: Jul 7, 2026

Cost-Efficient Transcriptomic-Based Drug Screening
06:40

Cost-Efficient Transcriptomic-Based Drug Screening

Published on: February 23, 2024

Robust-LongSAGE (RL-SAGE): an improved LongSAGE method for high-throughput transcriptome analysis.

Malali Gowda1, Guo-Liang Wang

  • 1Ohio State University, Columbus, Ohio, USA.

Methods in Molecular Biology (Clifton, N.J.)
|February 22, 2008
PubMed
Summary
This summary is machine-generated.

Robust-LongSAGE (RL-SAGE) enhances Serial Analysis of Gene Expression (SAGE) library construction. This improved method significantly increases cloning efficiency and insert size, enabling large-scale transcriptome analysis.

More Related Videos

Enrichment of mRNA and Bisulfite-mRNA Library Preparation for Next-Generation Sequencing
06:57

Enrichment of mRNA and Bisulfite-mRNA Library Preparation for Next-Generation Sequencing

Published on: July 7, 2023

Related Experiment Videos

Last Updated: Jul 7, 2026

Cost-Efficient Transcriptomic-Based Drug Screening
06:40

Cost-Efficient Transcriptomic-Based Drug Screening

Published on: February 23, 2024

Enrichment of mRNA and Bisulfite-mRNA Library Preparation for Next-Generation Sequencing
06:57

Enrichment of mRNA and Bisulfite-mRNA Library Preparation for Next-Generation Sequencing

Published on: July 7, 2023

Area of Science:

  • Molecular Biology
  • Genomics
  • Bioinformatics

Background:

  • Serial Analysis of Gene Expression (SAGE) is a valuable tool for transcriptome analysis.
  • Traditional SAGE library construction faces challenges like low cloning efficiency and small insert sizes.
  • These limitations hinder its routine application in many labs.

Purpose of the Study:

  • To improve the efficiency and reliability of SAGE library construction.
  • To develop a modified SAGE method suitable for high-throughput expression profiling.
  • To overcome the technical hurdles associated with standard LongSAGE.

Main Methods:

  • Development of Robust-LongSAGE (RL-SAGE), a modified LongSAGE library construction protocol.
  • Optimization of concatemer cloning and insert size.
  • Application of RL-SAGE for generating multiple gene expression libraries.

Main Results:

  • RL-SAGE significantly enhanced concatemer cloning efficiency and clone insert size compared to standard LongSAGE.
  • A library with over 150,000 clones can be generated using approximately 20 PCR reactions.
  • Successfully created 10 RL-SAGE libraries for rice, maize, and Magnaporthe grisea.

Conclusions:

  • RL-SAGE offers a robust and efficient method for large-scale transcriptome analysis.
  • The improved protocol addresses key limitations of previous SAGE techniques.
  • RL-SAGE facilitates routine gene expression profiling in diverse eukaryotic systems.