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An optimized protocol for rapid and sensitive application of southern (Northern) hybridization by using fluorescein labeling and detecting system

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Biotechnology Techniques

Abstract

An optimized complete protocol that produces consistent Southern hybridization results (RFLP) for as many as 240 samples in 36 h is presented. Signals can be detected from 0.1 μg genomic DNA. The protocol can be adapted to Northern hybridization by using 0.05 M NaOH as transfer buffer in a downward blotting method. Probes can be removed in stripping solution containing 2–5 mM EDTA. The protocol using this system has many advantages over the conventional radioactive method as safety, rapidness, sensitivity, and signal quality.

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References

  • Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K (1990) Current Protocols in Molecular Biology. New York: Greene and Wiley Interscience.

    Google Scholar 

  • Ingelbrecht IL, Mandelbaum CI, Mirkov TE (1998) Highly sensitive Northern hybridization using a rapid protocol for downward alkaline blotting of RNA. Bio Techniques 25: 420-423.

    Google Scholar 

  • Koetsier PA, Schorr J, Doerfler W (1993) A rapid optimized protocol for downward alkaline Southern blotting of DNA. Bio Techniques 2: 260-262.

    Google Scholar 

  • Lu YH, Merlino M, Isaac PG, Staccy J, Bernard M, Leroy P (1994) A comparative analysis between [32P] and digoxigenin-labelled single-copy probes for RFLP detection in wheat. Agronomie 14: 33-39.

    Google Scholar 

  • Neuhaus-Url G, Neuhaus G (1993) The use of the nonradioactive digoxigenin chemiluminescent technology for plant genomic Southern blot hybridizarion: a comparison with radioactivity. Transgenic Res. 2: 115-120.

    Google Scholar 

  • Zheng XY, Wolff DW, Baudracco-Arnas S, Pitrat M (1999) Development and utility of cleaved amplified polymorphic sequences (CAPS) and restriction fragment length polymorphisms (RFLP) linked to the Fom-2 Fusarium wilt resistance gene in melon (Cucumis melo L.). Theor. Appl. Genet. (in press).

  • Zheng XY, Wolff DW, Danin-Polig Y, Katzir N (1999) The association of ethylene production with postharvest fruit decay and DNA polymorphisms linked to ethylene production genes in melon (Cucumis melo L.) Theor. Appl. Genet. (in press).

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Authors and Affiliations

  1. Texas Agricultural Experiment Station, The Texas A & M University System, 2415 E. Highway 83, Weslaco, TX, 78596, USA

    X. Y. Zheng

  2. Sakata Seed America, Inc., P. O. Box 1118, Lehigh Acres, FL, 33970-1118, USA

    David W. Wolff

Authors
  1. X. Y. Zheng
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  2. David W. Wolff
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Zheng, X.Y., Wolff, D.W. An optimized protocol for rapid and sensitive application of southern (Northern) hybridization by using fluorescein labeling and detecting system. Biotechnology Techniques 13, 431–435 (1999). https://doi.org/10.1023/A:1008916402379

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  • DOI: https://doi.org/10.1023/A:1008916402379

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