Isolation of megabase-size DNA from sorghum and applications for physical mapping and bacterial and yeast artificial chromosome library construction

Sung Sick Woo, Vipin K. Rastogi, Hong Bin Zhang, Andrew H. Paterson, Keith F. Schertz, Rod A Wing

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

A method was developed for the isolation of megabase-size DNA from Sorghum bicolor. Sorghum protoplasts were isolated from young leaf tissue, embedded in an agarose matrix as microbeads or plugs, followed by cell lysis and protein degradation. The DNA prepared by this method was larger than 1 Mb in size and readily digestible with restriction enzymes. The DNA was shown to be suitable for physical mapping, and was successfully used for the construction of BAC and YAC libraries.

Original languageEnglish (US)
Pages (from-to)82-94
Number of pages13
JournalPlant Molecular Biology Reporter
Volume13
Issue number1
DOIs
StatePublished - Mar 1995
Externally publishedYes

Fingerprint

yeast artificial chromosomes
Yeast Artificial Chromosomes
Bacterial Artificial Chromosomes
bacterial artificial chromosomes
Sorghum
physical chromosome mapping
Chromosomes
Sorghum (Poaceae)
Yeast
DNA
Protoplasts
protein degradation
Microspheres
Sorghum bicolor
Sepharose
agarose
protoplasts
Proteolysis
Libraries
Tissue

Keywords

  • Megabase-size DNA
  • Physical mapping
  • Sorghum bicolor
  • YAC and BAC cloning

ASJC Scopus subject areas

  • Biochemistry
  • Biochemistry, Genetics and Molecular Biology(all)
  • Plant Science
  • Molecular Biology

Cite this

Isolation of megabase-size DNA from sorghum and applications for physical mapping and bacterial and yeast artificial chromosome library construction. / Woo, Sung Sick; Rastogi, Vipin K.; Zhang, Hong Bin; Paterson, Andrew H.; Schertz, Keith F.; Wing, Rod A.

In: Plant Molecular Biology Reporter, Vol. 13, No. 1, 03.1995, p. 82-94.

Research output: Contribution to journalArticle

Woo, Sung Sick ; Rastogi, Vipin K. ; Zhang, Hong Bin ; Paterson, Andrew H. ; Schertz, Keith F. ; Wing, Rod A. / Isolation of megabase-size DNA from sorghum and applications for physical mapping and bacterial and yeast artificial chromosome library construction. In: Plant Molecular Biology Reporter. 1995 ; Vol. 13, No. 1. pp. 82-94.
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