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The construction and characteristics of a BAC library for Cucumis sativus L. ‘B10’

Abstract

Cloning using bacterial artificial chromosomes (BACs) can yield high quality genomic libraries, which are used for the physical mapping, identification and isolation of genes, and for gene sequencing. A BAC genomic library was constructed from high molecular weight DNA (HMW DNA) obtained from nuclei of the cucumber (Cucumis sativus L. cv. Borszczagowski; B10 line). The DNA was digested with the HindIII restriction enzyme and ligated into the pCC1BAC vector. The library consists of 34,560 BAC clones with an average insert size of 135 kb, and 12.7x genome coverage. Screening the library for chloroplast and mitochondrial DNA content indicated an exceptionally low 0.26% contamination with chloroplast DNA and 0.3% with mitochondrial DNA.

Abbreviations

BAC:

bacterial artificial chromosome

BSA:

albumin from bovine serum

DAPI:

4′-6′-Diamidino-2-phenylindole dihydrochloride

HMW DNA:

high molecular weight DNA

LMP:

low melting agarose

PCR:

polymerase chain reaction

PFGE:

pulsed field gel electrophoresis

PVP:

polyvinylpyrrolidone

SCAR:

sequence-characterised amplified regions

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Correspondence to Wojciech Gutman.

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Gutman, W., Pawełkowicz, M., Woycicki, R. et al. The construction and characteristics of a BAC library for Cucumis sativus L. ‘B10’. Cell Mol Biol Lett 13, 74–91 (2008). https://doi.org/10.2478/s11658-007-0038-6

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Key words

  • Cucumis sativus L.
  • Bacterial artificial chromosome
  • High molecular weight DNA
  • Pulsed field gel electrophoresis