The development and application of automated gridding for efficient screening of yeast and bacterial ordered libraries.
An automated gridding procedure for the inoculation of yeast and bacterial clones in high-density arrays has been developed. A 96-pin inoculating tool compatible with the standard microtiter plate format and an eight-position tablet have been designed to fit the Biomek 1000 programmable robotic workstation (Beckman Instruments). The system is used to inoculate six copies of 80 x 120-mm filters representing a total of approximately 20,000 individual clones in approximately 3 h. High-density arrays of yeast artificial chromosome (YAC) and cosmid clones have been used for rapid large-scale hybridization screens of ordered libraries. In addition, an improved PCR library screening strategy has been developed using strips cut from the high-density arrays to prepare row and column DNA pools for PCR analysis. This strategy eliminates the final hybridization step and allows identification of a single clone by PCR in 2 days. The development of automated gridding technology will have a significant impact on the establishment of fully versatile screening of ordered library resources for genomic studies.
Bentley
D R
DR
Division of Medical and Molecular Genetics, United Medical School of Guy's Hospital, London, United Kingdom.
Todd
C
C
Collins
J
J
Holland
J
J
Dunham
I
I
Hassock
S
S
Bankier
A
A
Giannelli
F
F
D000818:Animals; D001322:Autoanalysis / Q000295:instrumentation* / Q000379:methods; D001419:Bacteria / Q000235:genetics*; D003001:Cloning, Molecular / Q000379:methods*; D003360:Cosmids; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D015342:DNA Probes; D015723:Gene Library; D006801:Humans; D010761:Phosphorus Radioisotopes; D012371:Robotics; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D013462:Sulfur Radioisotopes
D015342:DNA Probes; D010761:Phosphorus Radioisotopes; D013462:Sulfur Radioisotopes; D004247:DNA