Construction and characterisation of a yeast artificial chromosome library containing three haploid maize genome equivalents

Keith J. Edwards; Helen Thompson; David Edwards; Antoine de Saizieu; Caroline Sparks; John A. Thompson; Andrew J. Greenland; Mark Eyers; Wolfgang Schuch

doi:10.1007/BF00027351

Construction and characterisation of a yeast artificial chromosome library containing three haploid maize genome equivalents

Keith J. Edwards, Helen Thompson, David Edwards, Antoine de Saizieu, Caroline Sparks, John A. Thompson, Andrew J. Greenland, Mark Eyers, Wolfgang Schuch

Research output: Contribution to journal › Article › peer-review

67 Citations (Scopus)

Abstract

We have constructed a yeast artificial chromosome (YAC) library using high-molecular-weight DNA prepared from agarose-embedded leaf protoplasts of the maize inbred line UE95. This library contains 79 000 clones with an average insert size of 145 kb and should therefore represent approximately three haploid genome equivalents. The library is organised as an ordered array in duplicate microtitre plates. Forty-one pools of DNA from 1920 individual clones have been prepared for rapid screening of the library by the polymerase chain reaction (PCR). Using this approach, together with conventional colony hybridisation, we have been able to identify between one and eight positive clones for every probe used.

Original language	English
Pages (from-to)	299-308
Number of pages	10
Journal	Plant Molecular Biology
Volume	19
Issue number	2
DOIs	https://doi.org/10.1007/BF00027351
Publication status	Published - 1 May 1992
Externally published	Yes

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AU - de Saizieu, Antoine

AU - Sparks, Caroline

AU - Thompson, John A.

AU - Greenland, Andrew J.

AU - Eyers, Mark

AU - Schuch, Wolfgang

PY - 1992/5/1

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AB - We have constructed a yeast artificial chromosome (YAC) library using high-molecular-weight DNA prepared from agarose-embedded leaf protoplasts of the maize inbred line UE95. This library contains 79 000 clones with an average insert size of 145 kb and should therefore represent approximately three haploid genome equivalents. The library is organised as an ordered array in duplicate microtitre plates. Forty-one pools of DNA from 1920 individual clones have been prepared for rapid screening of the library by the polymerase chain reaction (PCR). Using this approach, together with conventional colony hybridisation, we have been able to identify between one and eight positive clones for every probe used.

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Construction and characterisation of a yeast artificial chromosome library containing three haploid maize genome equivalents

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