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A microassay for detection of DNA and RNA in small numbers of plant cells

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Summary

A microtechnique for the detection of DNA or RNA in small numbers of plant cells (1–50) has been developed using cauliflower mosaic virus (CaMV) infection of turnip as a model system. Both DNA and RNA extracted from 10 mesophyll protoplasts from CaMV-infected plants can be detected by hybridization using a radioactive probe made from cloned CaMV DNA (pCaMV10). No hybridization above background was detected in extracts of protoplasts from uninfected plants. At least 0.15 pg (11 000 molecules) of purified pCaMV10 DNA can be detected. This method is superior to existing ‘macro’ techniques for nucleic acid detection as smaller amounts of tissue are required and the detection is approximately 100-fold more sensitive. re]19850326 rv]19850530 ac]19850611

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References

  1. ThomasPS: Proc Natl Acad Sci USA 77:5201–5205, 1980.

    Google Scholar 

  2. WhiteBA, BancroftFC: J Biol Chem 257:8569–8572, 1982.

    Google Scholar 

  3. ManzariV, GalloRC, FranchiniG, WestinE, Ceccherini-NelliL, PopovicM, Wong-StaalF: Proc Natl Acad Sci USA 80:11–15, 1983.

    Google Scholar 

  4. HowarthAJ, GardnerRC, MessingJ, ShepherdRJ: Virology 112:678–685, 1981.

    Google Scholar 

  5. Maniatis T, Fritsch EF, Sambrook J: Molecular Cloning, Cold Spring Harbor Laboratory, 309 ff, 1982.

  6. GardnerRC, HowarthAJ, HahnP, Brown-LuediM, ShepherdRJ, MessingJ: Nucleic Acids Res 9:2871–2887, 1981.

    Google Scholar 

  7. ShewmakerCK, CatonJR, HouckCM, GardnerRC: Virology 140:281–288, 1985.

    Google Scholar 

  8. HullR, CoveySN: Nucleic Acids Res 11:1881–1895, 1983.

    Google Scholar 

  9. HullR, CoveySN: Trends in Biochem Sci 88:19–121, 1983.

    Google Scholar 

  10. EdstromJE: In: PrescottDM (ed) Methods in cell physiology. Academic Press, New York, 1964, pp 417–447.

    Google Scholar 

  11. ScalengheF, TurcoE, EdstromJE, PirrottaV, MelliM: Chromosoma (Berl) 82:205–216, 1981.

    Google Scholar 

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Authors and Affiliations

  1. Calgene, Inc., 1920 Fifth St., 95616, Davis, CA, U.S.A.

    Anne Crossway & Catherine M. Houck

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  1. Anne Crossway
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  2. Catherine M. Houck
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Crossway, A., Houck, C.M. A microassay for detection of DNA and RNA in small numbers of plant cells. Plant Mol Biol 5, 183–190 (1985). https://doi.org/10.1007/BF00015682

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  • DOI: https://doi.org/10.1007/BF00015682

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