title: Screening yeast artificial chromosome libraries with robot-aided automation.
abstract: Screening of collections of yeast artificial chromosomes utilizing the polymerase chain reaction (PCR) requires large numbers of reactions in parallel. Four steps were implemented to reduce the labor involved: (a) The number of initial samples for DNA extractions was decreased by compressing libraries up to 12-fold. (b) DNA extraction from yeast clones was robot assisted. (c) A BIOMEK 1000 station was adapted to pipette samples for PCR assays. (d) Sample preparation was integrated with a temperature cycler constructed to carry out up to 576 reactions in six 8 x 12-well trays. The implementation of these steps increases the number of reactions per person per day by an order of magnitude. In tests with X-chromosome-specific probes, the robot-aided screening recovered all of the clones detected by slower manual methods.
authors.affiliation: Washington University School of Medicine, Center for Genetics in Medicine, St. Louis, Missouri 63110.
authors.forename: D D
authors.initials: DD
authors.lastname: Sloan
authors.forename: M M
authors.initials: MM
authors.lastname: Blanchard
authors.forename: F W
authors.initials: FW
authors.lastname: Burough
authors.forename: V
authors.initials: V
authors.lastname: Nowotny
mesh_terms: D018244:Chromosomes, Artificial, Yeast; D004247:DNA / Q000302:isolation & purification; D005914:Globins / Q000235:genetics; D006801:Humans; D016133:Polymerase Chain Reaction / Q000295:instrumentation* / Q000379:methods; D012371:Robotics / Q000295:instrumentation*; D016324:Sequence Tagged Sites; D012984:Software; D014960:X Chromosome
chemical_list: D005914:Globins; D004247:DNA