Articles by Bernhard Michael Polzer in JoVE
Target Cell Pre-enrichment and Whole Genome Amplification for Single Cell Downstream Characterization Shukun Chen*1, Amin El-Heliebi*1, Julia Schmid1, Karl Kashofer2, Zbigniew T. Czyż3, Bernhard Michael Polzer3, Klaus Pantel4, Thomas Kroneis1,5, Peter Sedlmayr1 1Institute of Cell Biology, Histology and Embryology, Medical University of Graz, 2Institute of Pathology, Medical University of Graz, 3Fraunhofer Institute for Toxicology and Experimental Medicine ITEM, 4Department of Tumor Biology, University Medical Center Hamburg-Eppendorf, 5Sahlgrenska Cancer Center, University of Gothenburg This protocol is to recover and prepare rare target cells from a mixture with non-target background cells for molecular genetic characterization at the single-cell level. DNA quality is equal to non-treated single cells and allows for single-cell application (both screening based and targeted analysis).
Other articles by Bernhard Michael Polzer on PubMed
Principles of Whole-Genome Amplification Methods in Molecular Biology (Clifton, N.J.). | Pubmed ID: 26374306 Modern molecular biology relies on large amounts of high-quality genomic DNA. However, in a number of clinical or biological applications this requirement cannot be met, as starting material is either limited (e.g., preimplantation genetic diagnosis (PGD) or analysis of minimal residual cancer) or of insufficient quality (e.g., formalin-fixed paraffin-embedded tissue samples or forensics). As a consequence, in order to obtain sufficient amounts of material to analyze these demanding samples by state-of-the-art modern molecular assays, genomic DNA has to be amplified. This chapter summarizes available technologies for whole-genome amplification (WGA), bridging the last 25 years from the first developments to currently applied methods. We will especially elaborate on research application, as well as inherent advantages and limitations of various WGA technologies.
Laser Microdissection of FFPE Tissue Areas and Subsequent Whole Genome Amplification by Ampli1™ Methods in Molecular Biology (Clifton, N.J.). | Pubmed ID: 26374316 Laser microdissection (LMD) and whole genome amplification (WGA) are valuable tools to isolate, purify, and genetically analyze cancer cells from tissue sections. In this chapter, we describe a workflow for microdissecting small regions of interest from cancer tissue, i.e. formalin-fixed paraffin-embedded (FFPE) and cryo-conserved specimens, and subsequent whole genome amplification by a deterministic WGA approach (Ampli1™ WGA).