Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2012 Jan 26;7(2):328-38.
doi: 10.1038/nprot.2011.447.

Single-tube linear DNA amplification for genome-wide studies using a few thousand cells

Pattabhiraman Shankaranarayanan  1 , Marco-Antonio Mendoza-Parra, Wouter van Gool, Luisa M Trindade, Hinrich Gronemeyer
Affiliations

Single-tube linear DNA amplification for genome-wide studies using a few thousand cells

Pattabhiraman Shankaranarayanan et al. Nat Protoc. .

Abstract

Linear amplification of DNA (LinDA) by T7 polymerase is a versatile and robust method for generating sufficient amounts of DNA for genome-wide studies with minute amounts of cells. LinDA can be coupled to a great number of global profiling technologies. Indeed, chromatin immunoprecipitation coupled to massive parallel sequencing (ChIP-seq) has been achieved for transcription factors and epigenetic modification of chromatin histones with 1,000 to 5,000 cells. LinDA largely simplifies reChIP-seq experiments to monitor co-binding at chromatin target sites. The single-tube design of LinDA is ideal for handling ultrasmall amounts of DNA (<30 pg) and is compatible with automation. The actual hands-on working time is less than 6 h with one overnight reaction. The present protocol describes all materials and critical steps, and provides examples and controls for LinDA. Applications of LinDA for genome-wide analyses of biobank samples and for the study of chromatin conformation and nuclear architecture are in progress.

PubMed Disclaimer

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Nucleic Acids Res. 2008 Mar;36(4):e21 - PubMed
    1. Science. 2009 Oct 9;326(5950):289-93 - PubMed
    1. Nucleic Acids Res. 2008 Sep;36(16):e105 - PubMed
    1. Nature. 2010 Aug 12;466(7308):835-40 - PubMed
    1. Nat Rev Genet. 2009 Oct;10(10):669-80 - PubMed

Publication types

MeSH terms

LinkOut - more resources