Methods and Equipment


All analyses are PCR-based. For identification we mostly sequence a diagnostic fragment such as COI for insects (Brunner and Frey 2002). Where possible, this fragment is also used for population genetic analyses (Brunner et al. 2004). However, population genetics is in general studied using microsatellites (Brunner and Frey 2004, Pfunder and Frey 2005). In cases where mass-screening is required we develop real-time PCR assays that obviate the need for gel electrophoresis (Frey et al. 2004). This same technology is used where gene quantification is needed, such as in the quantitative assessment of heteroplasmy levels in weeds (Frey and Frey 2005). Microarray technology is being developed to simplify difficult diagnoses and multiplex analyses (Pfunder and Frey, 2004, Pfunder et al. 2004, Deyong et al. 2004). The technology is based on glass slides printed in-house with amino-linked oligonucleotides. Detection relies on fluorescent and gold labelling.


Our laboratory is fully equipped for PCR-based molecular work including sequencing, fragment analysis, cloning and microarray projects. We dispose of eight 96-well thermal cyclers (Perkin-Elmer, Techne) including one gradient cycler (Mastercycler gradient, Eppendorf), a pipetting robot (epMotion 5075, Eppendorf), a robot for magnetic separation (BioSprint 96, Qiagen), a 16-capillary sequencer and fragment analyzer (3130xl Genetic Analyzer, Applied Biosystems), real-time PCR machines (7500, Applied Biosystems), a DNA concentrator (5301, Eppendorf), equipment for DNA quantification (NanoDrop ND-1000 spectrophotometer, ND-3300 fluorospectrometer, Witec) and for gel electrophoresis and visualisation (UV light cabinet, AlphaInnoTech). For microarray analyses we dispose of an automated spotter for solid and split pin spotting (QArray Mini, Genetix), an automated slide hybridization station (SlidePro, GE-Amersham), and two slide readers for fluorescent signals (GenePix 4100A) and for silver and gold staining (HiLight Reader, Invitrogen).