Plant & Animal Genomes XI Conference

January 11-15, 2003
Town & Country Convention Center
San Diego, CA

Poster: High-throughput Methods
            

APPLICATION OF AUTOMATED COLONY PICKERS TO SEQUENCING TEMPLATE PREPARATION AND PCR SCREENING OF DNA LIBRARIES

Genetix Ltd, Queensway, New Milton, Hampshire, BH25 5NN, UK

Automated colony picking for the creation of microplate arrays of DNA clone libraries has become standard procedure in high-throughput genome sequencing laboratories. This is followed by the time-consuming and relatively expensive procedure of preparing plasmid DNA to use as a sequencing template or as template for PCR reactions in screening libraries for particular sequences. We have demonstrated that a colony picking robot can be used to pick colonies directly into TempliPhi (TM, Amersham Biosciences) reaction mix or into PCR master mix without cross contamination caused by carryover of cells or DNA on the robot pins. Carryover is eliminated using the appropriate pin washing and drying procedures, incorporating exposure of the pins to heat and light from a halogen lamp (patent pending). The combination of heat and UV irradiation serves to prevent DNA carryover to subsequent amplification steps. This means that the plasmid preparation step in genome sequencing can be eliminated reducing time and costs and increasing throughput.