Modified CTAB Procedure for DNA Isolation from Epiphytic Cacti of the Genera Hylocereus and Selenicereus (Cactaceae)

N. Tel-Zur, S. Abbo, D. Myslabodski, Y. Mizrahi

Research output: Contribution to journalArticlepeer-review

258 Scopus citations

Abstract

We present a simple protocol for DNA isolation from climbing cacti, genera Hylocereus and Selenicereus. The abundant polysaccharides present in Hylocereus and Selenicereus species interfere with DNA isolation, and DNA extracts, rich in polysaccharides, are poor templates for amplification using polymerase chain reaction (PCR). We used roots as the source tissue due to the lower viscosity of the extracts relative to that of other tissues. The extraction and isolation procedure we devised consists of the following steps: (1) three washes of ground tissue with the extraction buffer to remove the polysaccharides; (2) extraction with high-salt (4 M NaCl) cetyltrimethylammonium bromide (CTAB) buffer to remove the remaining polysaccharides; (3) removal of RNA by RNase; (4) phenol:chloroform extraction to remove proteins; (5) chloroform extraction to remove remaining phenols. The yields ranged from 10 to 20 μg DNA/g fresh roots. DNA samples prepared by our method were consistently amplifiable in the RAPD reaction and gave reproducible profiles.

Original languageEnglish
Pages (from-to)249-254
Number of pages6
JournalPlant Molecular Biology Reporter
Volume17
Issue number3
DOIs
StatePublished - 1 Jan 1999

Keywords

  • Cacti
  • DNA extraction
  • Hylocereus
  • Polysaccharides
  • Selenicereus

ASJC Scopus subject areas

  • Molecular Biology
  • Plant Science

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