The development and application of automated gridding for efficient screening of yeast and bacterial ordered libraries

D.R. Bentley, C. Todd, J. Collins, J. Holland, I. Dunham, S. Hassock, A. Bankier, F. Giannelli

Research output: Contribution to journalArticle

Abstract

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 ~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.
Original languageEnglish
Pages (from-to)534-541
Number of pages8
JournalGenomics
Volume12
Issue number3
DOIs
Publication statusPublished - Mar 1992

Keywords

  • article
  • automation
  • Bacteria
  • DNA library
  • molecular cloning
  • nonhuman
  • polymerase chain reaction
  • screening
  • yeast
  • yeast artificial chromosome

Fingerprint Dive into the research topics of 'The development and application of automated gridding for efficient screening of yeast and bacterial ordered libraries'. Together they form a unique fingerprint.

  • Cite this

    Bentley, D. R., Todd, C., Collins, J., Holland, J., Dunham, I., Hassock, S., Bankier, A., & Giannelli, F. (1992). The development and application of automated gridding for efficient screening of yeast and bacterial ordered libraries. Genomics, 12(3), 534-541. https://doi.org/10.1016/0888-7543(92)90445-X