STAR Protocols
Volume 2, Issue 4, 17 December 2021, 100991
Journal home page for STAR Protocols

Protocol
Global Run-On sequencing to measure nascent transcription in C. elegans

https://doi.org/10.1016/j.xpro.2021.100991Get rights and content
Under a Creative Commons license
open access

Highlights

  • Isolation of nuclei from C. elegans adults or embryos

  • Labeling of nascent transcripts with a biotinylated nucleotide

  • Three immunoprecipitation steps allow the efficient removal of the background

  • The library preparation strategy allows further removal of the background

Summary

Global Run-On sequencing (GRO-seq) is one of the most sensitive techniques to detect nascent transcription from RNA polymerase (Pol) at a genome-wide level. The protocol incorporates labeled ribonucleotides into nascent RNAs from Pol I, II, and III. We have adapted the GRO-seq protocol to the nematode Caenorhabditis elegans to measure transcription from embryos and adult worms. Here, we provide a detailed overview of the protocol highlighting the critical steps for generating successful libraries.

For complete details on the use and execution of this protocol, please refer to Quarato et al. (2021).

Subject areas

Cell separation/fractionation
Gene Expression
Genomics
Model Organisms
Molecular Biology
RNAseq
Sequencing

Data and code availability

Data are available upon reasonable request.

Cited by (0)

2

Technical contact

3

Lead contact