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Normalisation of cereal endosperm EST libraries for structural and functional genomic analysis

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Abstract

A simple method for the normalisation of cDNA libraries is presented. The method has been used to normalise barley and wheat endosperm libraries. Gene probes for transcripts present at different frequencies in the primary library were used to evaluate the effectiveness of the normalisation process.

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Abbreviations

BAC:

bacterial artificial chromosome

Cot:

initial DNA concentration in mol per litre times the re-association time in s

DPA:

days post-anthesis; d

ds-plasmid DNA:

double-stranded-plasmid DNA

EST:

expressed sequence tag

FAD3:

fatty acid desaturase 3

HAP:

hydroxyapatite

IPTG:

isopropylthio-β-D-galactoside

pfu:

plaque forming unit

SBEII:

starch branching enzyme II

ss-plasmid DNA:

single-stranded-plasmid DNA

SSI:

soluble starch synthase I

X-gal:

5-bromo-4-chloro-3-indolyl-β-D-galactoside

YAC:

yeast artificial chromosome

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Author notes

  1. Shahjahan Ali

    Present address: CSIRO Plant Industry, GPO Box 1600, 2601, Canberra, Australia

Authors and Affiliations

  1. CSIRO Plant Industry, GPO Box 1600, 2601, Canberra, Australia

    Bethwyn Holloway & William C. Taylor

Authors
  1. Shahjahan Ali
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  2. Bethwyn Holloway
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  3. William C. Taylor
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Correspondence to Shahjahan Ali.

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Ali, S., Holloway, B. & Taylor, W.C. Normalisation of cereal endosperm EST libraries for structural and functional genomic analysis. Plant Mol Biol Rep 18, 123–132 (2000). https://doi.org/10.1007/BF02824020

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