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RNA isolation from recalcitrant plant tissue

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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.

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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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Authors and Affiliations

  1. Intercollege Graduate Program in Genetics, The Pennsylvania State University, 16802, University Park, PA, USA

    David J. Schultz

  2. The Department of Horticulture, The Pennsylvania State University, 102 Tyson Building, 16802, University Park, PA, USA

    Richard Craig

  3. The Department of Entomology, The Pennsylvania State University, 501 ASI Building, 16802, University Park, PA, USA

    Diana L. Cox-Foster & Ralph O. Mumma

  4. The Department of Biology, The Pennsylvania State University, 208 Mueller Building, 16802, University Park, PA, USA

    June I. Medford

Authors
  1. David J. Schultz
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  2. Richard Craig
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  3. Diana L. Cox-Foster
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  4. Ralph O. Mumma
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  5. June I. Medford
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Schultz, D.J., Craig, R., Cox-Foster, D.L. et al. RNA isolation from recalcitrant plant tissue. Plant Mol Biol Rep 12, 310–316 (1994). https://doi.org/10.1007/BF02669273

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