Journal
/
Génétique
/
Transcription Start Site Mapping Using Super-low Input Carrier-CAGE
Journal JoVE
Génétique
This content is Free Access.
Journal JoVE Génétique
Transcription Start Site Mapping Using Super-low Input Carrier-CAGE

Transcription Start Site Mapping Using Super-low Input Carrier-CAGE

DOI:
10.3791/59805-v

•

06:59 min

•

June 26, 2019

•

, ,
1Institute of Clinical Sciences, Faculty of Medicine,Imperial College London, 2MRC London Institute of Medical Sciences, 3Sars International Centre for Marine Molecular Biology,University of Bergen

Chapitres

  • 00:04Titre
  • 01:04Preparation of DNA Templates
  • 01:38Reverse Transcription
  • 02:24Purification of cDNA:RNA Hybrids Using Magnetic Beads
  • 03:42Cap-trapping
  • 05:16Control of Degradation Level and Determination of the Number of PCR Cycles
  • 05:39Results: DNA Quality and Validation of SLIC-CAGE Libraries
  • 06:26Conclusion

Summary

Traduction automatique

Traduction automatique

Cap Analysis of Gene Expression (CAGE) is a method for genome-wide quantitative mapping of mRNA 5’ends to capture RNA polymerase II transcription start sites at a single-nucleotide resolution. This work describes a low-input (SLIC-CAGE) protocol for generation of high-quality libraries using nanogram-amounts of total RNA.

Tags

Transcription Start Site MappingSLIC-CAGECarrier-CAGERNA Polymerase IIGene RegulationCore PromoterEnhancer DiscoveryDiagnostic MarkersEmbryonic DevelopmentReverse TranscriptionSPRI Magnetic Beads

Article

Embed

AJOUTER À LA PLAYLIST

Usage Statistics

Vidéos Connexes

check_url/fr/59805?article_type=v

In Vitro Transcription Assays and Their Application in Drug Discovery
check_url/fr/59805?article_type=v

Mapping RNA-RNA Interactions Globally Using Biotinylated Psoralen
check_url/fr/59805?article_type=v

Analysis of Termination of Transcription Using BrUTP-strand-specific Transcription Run-on (TRO) Approach
check_url/fr/59805?article_type=v

Bidirectional Retroviral Integration Site PCR Methodology and Quantitative Data Analysis Workflow
check_url/fr/59805?article_type=v

Retroviral Scanning: Mapping MLV Integration Sites to Define Cell-specific Regulatory Regions
check_url/fr/59805?article_type=v

Real-time Analysis of Transcription Factor Binding, Transcription, Translation, and Turnover to Display Global Events During Cellular Activation
check_url/fr/59805?article_type=v

Ultralow Input Genome Sequencing Library Preparation from a Single Tardigrade Specimen
check_url/fr/59805?article_type=v

Genome-wide Surveillance of Transcription Errors in Eukaryotic Organisms
check_url/fr/59805?article_type=v

Enhanced Yeast One-hybrid Screens To Identify Transcription Factor Binding To Human DNA Sequences
check_url/fr/59805?article_type=v

Discrimintion and Mapping of the Primary and Processed Transcripts in Maize Mitochondrion Using a Circular RT-PCR-based Strategy

Read Article