XACT-seq: A photocrosslinking-based technique for detection of the RNA polymerase active-center position relative to DNA in Escherichia coli

• Main Authors: Chirangini Pukhrambam, Irina O. Vvedenskaya, Bryce E. Nickels
• Format: Article
• Language: English
• Published: Elsevier 2021-12-01
• Series: STAR Protocols
• Subjects: Molecular Biology; Gene Expression; Protein Biochemistry
• Online Access: http://www.sciencedirect.com/science/article/pii/S2666166721005645
• ISSN: 2666-1667

Summary: XACT-seq (“crosslink between active-center and template sequencing”) is a technique for high-throughput, single-nucleotide resolution mapping of RNA polymerase (RNAP) active-center positions relative to the DNA template. XACT-seq overcomes limitations of approaches that rely on analysis of the RNA 3′ end (e.g., native elongating transcript sequencing) or that report RNAP positions with low resolution (e.g., ChIP-seq and ChIP-exo). XACT-seq can be used to map RNAP active-center positions in transcription initiation complexes, initially transcribing complexes, and transcription elongation complexes.For complete details on the use and execution of this protocol, please refer to Winkelman et al. (2020).