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In-cell RT-PCR in a single, detached plant cell

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Abstract

A procedure is presented for the detection of mRNA in a single root border cell of pea seedlings using RT-PCR (reverse transcription-polymerase chain reaction) and direct incorporation of digoxigenin-dUTP. It includes cell preservation with aldehyde. Signal is detected using a conjugate of anti-DIG and alkaline phosphatase.

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Abbreviations

DIG:

digoxigenin

DEPC:

diethylpyrocarbonate

DW:

distilled and deionized water

NBT:

nitroblue tetrazolium

PCR:

polymerase chain reaction

RT-PCR:

reverse transcription-polymerase chain reaction

SB:

staining buffer

SSB:

stain-stop buffer

X-PHOS:

5-bromo-4-chloro-3-indolyl phosphate

References

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Authors and Affiliations

  1. Departments of Plant Pathology and Molecular & Cellular Biology, University of Arizona, 85721, Tucson, AZ, USA

    Ho-Hyung Woo, Lindy A. Brigham & Martha C. Hawes

Authors
  1. Ho-Hyung Woo
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  2. Lindy A. Brigham
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  3. Martha C. Hawes
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Correspondence to Ho-Hyung Woo.

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Woo, HH., Brigham, L.A. & Hawes, M.C. In-cell RT-PCR in a single, detached plant cell. Plant Mol Biol Rep 13, 355–362 (1995). https://doi.org/10.1007/BF02669191

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  • DOI: https://doi.org/10.1007/BF02669191

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