Whole Genome Amplification by Isothermal Multiple Strand Displacement Using Phi29 DNA Polymerase

Thomas Kroneis; Amin El-Heliebi

doi:10.1007/978-1-4939-2990-0_8

Whole Genome Amplification by Isothermal Multiple Strand Displacement Using Phi29 DNA Polymerase

Methods Mol Biol. 2015:1347:111-7. doi: 10.1007/978-1-4939-2990-0_8.

Authors

Thomas Kroneis^{1

2}, Amin El-Heliebi³

Affiliations

¹ Research Unit for Single Cell Analysis, Institute of Cell Biology, Histology & Embryology, Medical University of Graz, Harrachgasse 21, Graz, 8010, Austria. thomas.kroneis@medunigraz.at.
² Sahlgrenska Cancer Center, University of Gothenburg, Gothenburg, Sweden. thomas.kroneis@medunigraz.at.
³ Research Unit for Single Cell Analysis, Institute of Cell Biology, Histology & Embryology, Medical University of Graz, Harrachgasse 21, Graz, 8010, Austria.

PMID: 26374313
DOI: 10.1007/978-1-4939-2990-0_8

Abstract

The here described method of isothermal whole genome amplification (iWGA) uses a Phi29 DNA polymerase-based kit (Illustra GenomiPhi V2 DNA Amplification Kit) that amplifies minute quantities of DNA by multiple strand displacement upon random hexamer primer binding. Starting from genomic DNA or single cells this amplification yields up to 5 μg of iWGA product with fragment lengths of 10 kb and longer. As this amplification lacks the need of fragmenting DNA, its products are well suited for many downstream applications (e.g. sequencing and DNA profiling). On the contrary, degraded DNA samples are not supported by the nature of the amplification and are not well suited.

Keywords: Isothermal whole genome amplification; Multiple strand displacement; Phi29 DNA polymerase; Single-cell analysis.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Bacteriophages / enzymology*
DNA-Directed DNA Polymerase*
Genome*
Genomics / methods*
Nucleic Acid Amplification Techniques*
Reagent Kits, Diagnostic
Single-Cell Analysis / methods

Substances

Reagent Kits, Diagnostic
DNA-Directed DNA Polymerase