A transcription-based approach to purify R-loop-containing plasmid DNA templates in vitro Journal Article

   


Authors: Kumar, C.; Remus, D.
Article Title: A transcription-based approach to purify R-loop-containing plasmid DNA templates in vitro
Abstract: To study the direct effects of R-loops on DNA replication and other DNA-templated processes in vitro, R-loop-containing DNA templates need to be prepared efficiently and to near homogeneity. Here, we describe a simple transcription-based approach to form R-loops on plasmid DNA templates in vitro. We detail steps to transcribe a DNA sequence element with a high propensity to form co-transcriptional R-loops using T7 RNA polymerase. We describe nucleolytic digestion of free RNA, deproteinization, and repurification of R-loop-containing templates via gel filtration. For complete details on the use and execution of this protocol, please refer to Kumar et al.1 © 2022 The Authors
Keywords: molecular biology; protein biochemistry
Journal Title: STAR Protocols
Volume: 4
Issue: 1
ISSN: 2666-1667
Publisher: Cell Press  
Date Published: 2023-03-17
Start Page: 101937
Language: English
DOI: 10.1016/j.xpro.2022.101937
PROVIDER: scopus
PMCID: PMC9758483
PUBMED: 36520635
DOI/URL:

https://www.scopus.com/inward/record.uri?eid=2-s2.0-85144854834&doi=10.1016%2fj.xpro.2022.101937&partnerID=40&md5=2f4a053fac254913fcdda1374d64219a
Notes: Article -- MSK Cancer Center Support Grant (P30 CA008748) acknowledged in PubMed and PDF -- MSK corresponding author is Dirk Remus -- Export Date: 1 February 2023 -- Source: Scopus
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MSK Authors
  1. Dirk Remus
    21 Remus
  2. Charanya Kumar
    8 Kumar
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