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

Fingerprint

Libraries
Clone Cells
Yeasts
Polymerase Chain Reaction
Yeast Artificial Chromosomes
Cosmids
Genomic Library
Robotics
Tablets
Technology
DNA

Keywords

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

Cite this

The development and application of automated gridding for efficient screening of yeast and bacterial ordered libraries. / Bentley, D.R.; Todd, C.; Collins, J.; Holland, J.; Dunham, I.; Hassock, S.; Bankier, A.; Giannelli, F.

In: Genomics, Vol. 12, No. 3, 03.1992, p. 534-541.

Research output: Contribution to journalArticle

Bentley, DR, 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, vol. 12, no. 3, pp. 534-541. https://doi.org/10.1016/0888-7543(92)90445-X
Bentley, D.R. ; Todd, C. ; Collins, J. ; Holland, J. ; Dunham, I. ; Hassock, S. ; Bankier, A. ; Giannelli, F. / The development and application of automated gridding for efficient screening of yeast and bacterial ordered libraries. In: Genomics. 1992 ; Vol. 12, No. 3. pp. 534-541.
@article{ee4b8592c45e49cb95d2c86ac03a83b6,
title = "The development and application of automated gridding for efficient screening of yeast and bacterial ordered libraries",
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.",
keywords = "article, automation, Bacteria, DNA library, molecular cloning, nonhuman, polymerase chain reaction, screening, yeast, yeast artificial chromosome",
author = "D.R. Bentley and C. Todd and J. Collins and J. Holland and I. Dunham and S. Hassock and A. Bankier and F. Giannelli",
year = "1992",
month = "3",
doi = "10.1016/0888-7543(92)90445-X",
language = "English",
volume = "12",
pages = "534--541",
journal = "Genomics",
issn = "0888-7543",
publisher = "Elsevier Academic Press Inc",
number = "3",

}

TY - JOUR

T1 - The development and application of automated gridding for efficient screening of yeast and bacterial ordered libraries

AU - Bentley, D.R.

AU - Todd, C.

AU - Collins, J.

AU - Holland, J.

AU - Dunham, I.

AU - Hassock, S.

AU - Bankier, A.

AU - Giannelli, F.

PY - 1992/3

Y1 - 1992/3

N2 - 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.

AB - 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.

KW - article

KW - automation

KW - Bacteria

KW - DNA library

KW - molecular cloning

KW - nonhuman

KW - polymerase chain reaction

KW - screening

KW - yeast

KW - yeast artificial chromosome

UR - http://dx.doi.org/10.1016/0888-7543(92)90445-X

U2 - 10.1016/0888-7543(92)90445-X

DO - 10.1016/0888-7543(92)90445-X

M3 - Article

VL - 12

SP - 534

EP - 541

JO - Genomics

JF - Genomics

SN - 0888-7543

IS - 3

ER -