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Plant Molecular Biology Reporter

, Volume 12, Issue 4, pp 310–316 | Cite as

RNA isolation from recalcitrant plant tissue

  • David J. Schultz
  • Richard Craig
  • Diana L. Cox-Foster
  • Ralph O. Mumma
  • June I. Medford
Commentary

Abstract

The isolation of high-quality RNA from various tissues (leaves, pedicels, glandular trichomes) of garden geranium (Pelargonium xhortorum) using various published methods is difficult due to numerous oxidizing compounds. A new RNA extraction method was developed through the combination and modification of two separate procedures (Rochester et al., 1986; Manning 1991). In addition to geranium tissues, this method is successful when used with other recalcitrant tissues such as mature needles of white pine (Pinus strobus) and mature leaves of poinsettia (Euphorbia pulcherrima). RNA quality was judged by spectrophotometric readings, denaturing agarose gels, and successful reverse transcription.

Key Words

RNA isolation geranium glandular trichomes recalcitrant plant tissue 

Abbreviations

CTAB

cetyltrimethylammonium bromide

EGTA

ethylene glycol-bis (β-aminoethyl ether) N, N, N-, N-tetraacetic acid

PVPP

polyvinylpolypyrrolidone

2-BE

ethylene glycol monobutylether

DEPC

diethylpyrocarbonate

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Copyright information

© Kluwer Academic Publishers 1994

Authors and Affiliations

  • David J. Schultz
    • 1
  • Richard Craig
    • 2
  • Diana L. Cox-Foster
    • 3
  • Ralph O. Mumma
    • 3
  • June I. Medford
    • 4
  1. 1.Intercollege Graduate Program in GeneticsThe Pennsylvania State UniversityUniversity ParkUSA
  2. 2.The Department of HorticultureThe Pennsylvania State UniversityUniversity ParkUSA
  3. 3.The Department of EntomologyThe Pennsylvania State UniversityUniversity ParkUSA
  4. 4.The Department of BiologyThe Pennsylvania State UniversityUniversity ParkUSA

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