Skip to main content
SpringerLink
Log in

PCR-Based Assays for Strand-Specific Measurement of DNA Damage and Repair II

Single-Strand Ligation-PCR

  • Protocol
DNA Repair Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 113))

  • 933 Accesses

Abstract

The previous chapter describes strand-specific quantitative QPCR (ss-QPCR), which enables DNA damage and repair in short gene regions (∬300 and upward) to be measured in a strand-specific manner. This chapter describes single-strand ligation PCR (sslig-PCR), which extends the previous method and allows damage and repair to be studied at the single-nucleotide level in single-copy genes in mammalian cells. The importance of this type of measurement is shown by the demonstration of a cell-specific adduct formed by the anticancer drug cisplatin, which is not formed when extracted DNA is treated in vitro (1).

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 129.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 169.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Grimaldi, K. A., McAdam, S. R., Souhami, R. L., and Hartley, J. A. (1994) DNA damage by anti-cancer agents resolved at the nucleotide level of a single copy gene: evidence for a novel binding site for cisplatin in cells. Nucleic Acids Res. 22, 2311–2317.

    Article  PubMed  CAS  Google Scholar 

  2. Pfeifer, G. P., Steigerwald, S. D., Mueller, P. R., Wold, B., Riggs, and Riggs, A. D. (1989) Genomic sequencing and methylation analysis by ligation mediated PCR. Science 246, 810–813.

    Article  PubMed  CAS  Google Scholar 

  3. Tessier, D., Brousseau, R., and Vernet, T. (1986) Ligation of single stranded oligodeoxyribonucleotides by T4 RNA ligase. Anal. Biochem. 158, 171–178.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. CRC Drug-DNA Interactions Research Group, Department of Oncology, University College London Medical School, London, UK

    Keith A. Grimaldi, Simon R. McAdam & John A. Hartley

Authors
  1. Keith A. Grimaldi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Simon R. McAdam
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. John A. Hartley
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. University of Dundee, Dundee, UK

    Daryl S. Henderson

Rights and permissions

Reprints and permissions

Copyright information

© 1999 Humana Press Inc.

About this protocol

Cite this protocol

Grimaldi, K.A., McAdam, S.R., Hartley, J.A. (1999). PCR-Based Assays for Strand-Specific Measurement of DNA Damage and Repair II. In: Henderson, D.S. (eds) DNA Repair Protocols. Methods in Molecular Biology™, vol 113. Humana Press. https://doi.org/10.1385/1-59259-675-4:241

Download citation

  • DOI: https://doi.org/10.1385/1-59259-675-4:241

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-802-8

  • Online ISBN: 978-1-59259-675-1

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation