Global Run-on Sequencing

GRO-Seq maps the binding sites of transcriptionally active RNA polymerase II (RNAPII) (Core et al., 2008). In this method, active RNAPII is allowed to run on in the presence of 5-bromouridine 5′-triphosphate (Br-UTP). RNAs are hydrolyzed and purified using beads coated with antibodies to 5-bromo-2-deoxyuridine (BrdU). After cap removal and end repair, the eluted RNA is reverse-transcribed to cDNA. Deep sequencing of the cDNA identifies RNAs that are actively transcribed by RNAPII.


  • Maps position of transcriptionally engaged RNA polymerases
  • Determines relative activity of transcription sites
  • Detects sense and antisense transcription
  • Detects transcription anywhere on the genome
  • No prior knowledge of transcription sites is needed
  • Provides robust coverage of enhancer- and promoter-associated RNAs (Melnik et al., 2016)


  • Limited to cell cultures and other artificial systems, due to the requirement for incubation in the presence of labeled nucleotides
  • Artifacts may be introduced during the preparation of the nuclei (Adelman et al., 2012)
  • New initiation events may occur during the run-on step
  • Physical impediments may block the polymerases
  • Resolution is only 30_100 nt (Nojima et al., 2016)
  • Requires nascent RNAs of at least 18 nt (Mayer et al., 2016).


Illumina Library prep and Array Kit Selector


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