Plant Molecular Biology Reporter

, Volume 10, Issue 3, pp 273–284

Amplification and sequencing of 16/18S rDNA from gel-purified total plant DNA

  • Carol Bult
  • Mari Källersjö
  • Youngbae Suh
Protocol

DOI: 10.1007/BF02668360

Cite this article as:
Bult, C., Källersjö, M. & Suh, Y. Plant Mol Biol Rep (1992) 10: 273. doi:10.1007/BF02668360

Abstract

We describe here methods and strategies for amplifying and sequencing the genes encoding the small subunits (16/18S) of nuclear and chloroplast ribosomal DNA (rDNA) from total plant DNA. These methods were developed in response to technical difficulties we encountered in our molecular systematic work with members of various plant families. These protocols have proved useful when the amount of tissue available for study is limited and when the tissues have high concentrations of undesirable secondary metabolites which are often co-isolated with nucleic acids.

Key words

DNA extraction gel purification PCR amplification double-stranded DNA sequencing rDNA nuclear chloroplast 

Abbreviations

dNTP

deoxynucleotide triphosphate

ddNTP

dideoxynucleotide triphosphate

rDNA

ribosomal DNA

CTAB

hexadecyltrimethylammonium bromide

Copyright information

© Kluwer Academic Publishers 1992

Authors and Affiliations

  • Carol Bult
    • 1
  • Mari Källersjö
    • 1
  • Youngbae Suh
    • 1
  1. 1.Laboratory of Molecular Systematics Smithsonian InstitutionNational Museum of Natural HistoryWashington, D.C.USA

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