Skip Navigation

This Article
Right arrow Full Text Freely available
Right arrow Print PDF (208K) Freely available
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrowRequest Permissions
Right arrow Commercial Re-use Guidelines
for Open Access NAR Content
Google Scholar
Right arrow Articles by Diggle, C. P.
Right arrow Articles by Kiltie, A. E.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Diggle, C. P.
Right arrow Articles by Kiltie, A. E.
Related Collections
Right arrow Repair
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Nucleic Acids Research, 2003, Vol. 31, No. 15 e83
© 2003 Oxford University Press

Development of a rapid, small-scale DNA repair assay for use on clinical samples

Christine P. Diggle, Johanne Bentley and Anne E. Kiltie*

Cancer Research UK Clinical Centre, St James’s University Hospital, Leeds LS9 7TF, UK

*To whom correspondence should be addressed. Tel: +44 113 206 4908; Fax: +44 113 242 9886; Email: anne.kiltie{at}cancer.org.uk

Double-strand breaks (DSBs) are the most lethal form of DNA damage. They can be repaired by one of two pathways, homologous recombination and non-homologous end joining (NHEJ). A NHEJ assay has previously been reported which measures joining using cell-free extracts and a linearised plasmid as DNA substrate. This assay was designed for 3 x 109 cells grown in vitro and utilised radioactively labelled substrate. We have scaled down the method to use smaller cell numbers in a variety of cell lines. Altering the cellular extraction procedure decreased background DNA contamination. The cleaner preparations allowed us to use SYBR Green I staining to identify joined products, which was as sensitive as 32P-end-labelled DNA. NHEJ was found in established tumour cell lines from different originating tissues, though actual levels and fidelity of repair differed. This method also allowed end joining to be assessed in clinical specimens (human blood, brain and bladder tumours) within 24 h of receiving samples. The application of this method will allow investigation of the role of DSB DNA repair pathways in human tumours.


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?


This article has been cited by other articles:


Home page
 CarcinogenesisHome page
N. Machella, M. B. Terry, J. Zipprich, I. Gurvich, Y. Liao, R. T. Senie, D. O. Kennedy, and R. M. Santella
Double-strand breaks repair in lymphoblastoid cell lines from sisters discordant for breast cancer from the New York site of the BCFR
Carcinogenesis, July 1, 2008; 29(7): 1367 - 1372.
[Abstract] [Full Text] [PDF]

Home page
 Clin. Chem.Home page
E. C. Timmermans, P. Tebas, J. P.N. Ruiter, R. J.A. Wanders, A. de Ronde, and M. P. de Baar
Real-Time Nucleic Acid Sequence-Based Amplification Assay to Quantify Changes in Mitochondrial DNA Concentrations in Cell Cultures and Blood Cells from HIV-Infected Patients Receiving Antiviral Therapy
Clin. Chem., June 1, 2006; 52(6): 979 - 987.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
C. P. Diggle, J. Bentley, M. A. Knowles, and A. E. Kiltie
Inhibition of double-strand break non-homologous end-joining by cisplatin adducts in human cell extracts
Nucleic Acids Res., May 4, 2005; 33(8): 2531 - 2539.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
J. Bentley, C. P. Diggle, P. Harnden, M. A. Knowles, and A. E. Kiltie
DNA double strand break repair in human bladder cancer is error prone and involves microhomology-associated end-joining
Nucleic Acids Res., October 5, 2004; 32(17): 5249 - 5259.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
H. Zipper, H. Brunner, J. Bernhagen, and F. Vitzthum
Investigations on DNA intercalation and surface binding by SYBR Green I, its structure determination and methodological implications
Nucleic Acids Res., July 12, 2004; 32(12): e103 - e103.
[Abstract] [Full Text] [PDF]


Disclaimer: Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.