@article{45430b6052e1463aaa32f1243e565bde,
title = "Small-scale extracts for the study of nucleotide excision repair and non-homologous end joining",
abstract = "The repair of DNA by nucleotide excision repair (NER) and non-homologous end joining (NHEJ) is essential for maintenance of genomic integrity and cell viability. Examination of NHEJ and NER in vitro using cell-free extracts has led to a deeper understanding of the biochemical mechanisms that underlie these processes. Current methods for production of whole-cell extracts (WCEs) to investigate NER and NHEJ start with one or more liters of culture containing 1-5 × 109 cells. Here, we describe a small-scale method for production of WCE that can be used to study NER. We also describe a rapid, small-scale method for the preparation of WCE that can be used in the study of NHEJ. These methods require less time, 20- to 1000-fold fewer cells than large-scale extracts, facilitate examination of numerous samples and are ideal for such applications as the study of host-virus interactions and analysis of mutant cell lines.",
author = "Smeaton, {Michael B.} and Miller, {Paul S.} and Gary Ketner and Hanakahi, {Les A.}",
note = "Funding Information: We thank Joyce Cheung, Timra Gilson, Brenda Salerno, Sumithra Jayaram, B.T. Rantipole and Bill Russ for many thoughtful discussions. We thank Drs Joyce Reardon and Aziz Sancar for helpful advice on preparing NER extracts; Ms Maggie Wear for assistance in oligonucleotide synthesis; and Dr Michael Seidman for generously providing CHO AA8 and CHO UV5 cell lines. UV41 cells were obtained from American type culture collection under grant number CA082785. HeLa cells were obtained from the National Cell Culture Center (Minneapolis, MN, USA), supported under grant number U42 RR05991 from the National Center for Research Resources, NIH. This work was supported by the National Institutes of Health (NIH) grants GM070639-1 (to L.A.H.), 5R01CA082127 (to G.K.) and CA082785 (to P.S.M.), and by the Johns Hopkins University Bloomberg School of Public Health Faculty Research Initiatives Fund (to L.A.H. and G.K.). Funding to pay the Open Access publication charges for this article was provided by National Institutes of Health (NIH) grants GM070639-1.",
year = "2007",
month = dec,
doi = "10.1093/nar/gkm974",
language = "English (US)",
volume = "35",
journal = "Nucleic acids research",
issn = "0305-1048",
publisher = "Oxford University Press",
number = "22",
}