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Transcription Start Site Mapping Using Super-low Input Carrier-CAGE
JoVE Journal
Genetics
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JoVE Journal Genetics
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

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06:59 min

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June 26, 2019

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, ,
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

Chapters

  • 00:04Title
  • 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

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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

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