Screening yeast artificial chromosome libraries with robot-aided automation.
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.
Sloan
D D
DD
Washington University School of Medicine, Center for Genetics in Medicine, St. Louis, Missouri 63110.
Blanchard
M M
MM
Burough
F W
FW
Nowotny
V
V
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
D005914:Globins; D004247:DNA