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PMID: 1559704 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

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

Genomics ·Vol. 12 ·No. 3 ·1992-03-00 ·Pages 534-41

Bentley DR, Todd C, Collins J, Holland J, Dunham I, Hassock S, Bankier A, Giannelli F

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 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.

MeSH Terms
Animals Autoanalysis/instrumentation,methods Bacteria/genetics Cloning, Molecular/methods Cosmids DNA/genetics,isolation & purification DNA Probes Gene Library Humans Phosphorus Radioisotopes Robotics Saccharomyces cerevisiae/genetics Sulfur Radioisotopes
Chemicals
DNA Probes Phosphorus Radioisotopes Sulfur Radioisotopes DNA
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Bentley D R
Division of Medical and Molecular Genetics, United Medical School of Guy's Hospital, London, United Kingdom.
Todd C
Collins J
Holland J
Dunham I
Hassock S
Bankier A
Giannelli F
Article Info
Journal
Genomics
Abbr.
Genomics
ISSN
0888-7543
Published
1992-03-00
Pages
534-41
Language
English
Region
United States
NLM ID
8800135
Subset
IM
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