title: Low copy number DNA profiling from isolated sperm using the aureka®-micromanipulation system.
abstract: A new cell isolation technique linked to the aureka® micromanipulation system (aureka®) was used to pick sperm from mixed samples containing sperm and epithelial cells. Both cell types were stained using the HY-LITER™ high-resolution, fluorescent staining kit. To isolate a single sperm of interest under a fluorescent microscope, a specific microsphere picking technique was used. This sensitive and reliable cell identification and isolation technique enables low-copy-number (LCN) DNA profiling, as few as 20 sperm are sufficient for obtaining a full short tandem repeat (STR) profile without any allelic drop out. The presented protocol covers the whole workflow, from sample staining and cell pick up to STR analysis.
authors.affiliation: Galantos Genetics GmbH, Johann-Joachim-Becher-Weg 30a, 55128 Mainz, Germany. cora.schneider@galantos.eu
authors.forename: C
authors.initials: C
authors.lastname: Schneider
authors.forename: U
authors.initials: U
authors.lastname: Müller
authors.forename: R
authors.initials: R
authors.lastname: Kilper
authors.forename: B
authors.initials: B
authors.lastname: Siebertz
mesh_terms: D002469:Cell Separation / Q000379:methods*; D004247:DNA / Q000032:analysis* / Q000235:genetics; D016172:DNA Fingerprinting / Q000379:methods*; D019075:Electrophoresis, Capillary; D004847:Epithelial Cells / Q000737:chemistry; D005260:Female; D006801:Humans; D008297:Male; D018895:Microsatellite Repeats; D008856:Microscopy, Fluorescence; D016133:Polymerase Chain Reaction; D013094:Spermatozoa / Q000737:chemistry*; D013194:Staining and Labeling
chemical_list: D004247:DNA