Using Multiplex PCR for Assessing the Quality of Whole Genome Amplified DNA

Amin El-Heliebi; Shukun Chen; Thomas Kroneis

doi:10.1007/978-1-4939-2990-0_9

Using Multiplex PCR for Assessing the Quality of Whole Genome Amplified DNA

Methods Mol Biol. 2015:1347:119-28. doi: 10.1007/978-1-4939-2990-0_9.

Authors

Amin El-Heliebi¹, Shukun Chen¹, Thomas Kroneis^{2

3}

Affiliations

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

PMID: 26374314
DOI: 10.1007/978-1-4939-2990-0_9

Abstract

This chapter describes a simple and inexpensive multiplex PCR-based method to assess the quality of whole genome amplification (WGA) products generated from heat-induced random fragmented DNA. A set of four primer pairs is used to amplify DNA sequences of WGA products in and downstream of GAPDH gene in yielding 100, 200, 300, and 400 bp fragments. PCR products are analyzed by agarose gel electrophoresis and the respective WGA quality is classified according to the number of obtained PCR bands. WGA products that yield three or four PCR bands are considered to be of high quality and yield good results when analyzed by means of array comparative genome hybridization (CGH).

Keywords: Quality control PCR; Single-cell analysis; Whole genome amplification.

MeSH terms

DNA*
Genome*
Genomics / methods
Genomics / standards
Multiplex Polymerase Chain Reaction / methods*
Nucleic Acid Amplification Techniques*
Quality Control*
Single-Cell Analysis / methods
Single-Cell Analysis / standards

Substances

DNA