Plant Molecular Biology Reporter

, Volume 11, Issue 1, pp 54–59 | Cite as

A protocol for non-radioactive DNA labelling and detection in the RFLP analysis of rice and tomato using single-copy probes

  • Olivier Panaud
  • Gerard Magpantay
  • Susan McCouch
Protocols

Abstract

An improved protocol for non-radioactive labelling and detection of rice and tomato DNA is described. The procedure includes the use of polymerase chain reaction for the incorporation of digoxigenin-dUTP in the DNA molecule and the use of a chemiluminescent compound (AMPPD) for the signal detection.

Key Words

RFLP rice tomato digoxigenin non-radioactive DNA labelling PCR 

Abbreviations

AP

alkaline phosphatase

dig

digoxigen

PCR

polymerase chain reaction

RFLP

restriction fragment length polymorphism

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References

  1. Allefs, J.J.H.M., E.M.J. Salentjin, F.A. Krens, G.J.A. Rouwental. 1990. Optimization of non-radioactive Southern blot hybridization: single copy detection and reuse of blots. Nucleic Acids Research 18:3099–3100.PubMedCrossRefGoogle Scholar
  2. Brar, D.S., B.G. de los Reyes, O. Panaud, A. Sanchez, and G.S. Khush. 1991. Genetic mapping in rice using isozyme and RFLP markers.Rice Genetics II, International Rice Research Institute, pp 137–146Google Scholar
  3. Ishii, T., O. Panaud, D.S. Brar, and G.S. Khush. 1990. Use of non-radioactive, digoxigenin-labeled DNA probes for RFLP analysis in rice. Plant Mol. Biol. Reptr. 8(3):167–171.Google Scholar
  4. Kreike, C.M., J.R.E. de Koning, F.A. Krens. 1990. Non-radioactive detection of single-copy DNA-DNA hybrids. Plant Mol. Biol. Reptr. 8(3):172–179.CrossRefGoogle Scholar
  5. Lanzillo, J. 1990. Preparation of digoxigenin-labeled probes by the polymerase chain reaction. BioTechniques 8:621–622.Google Scholar

Copyright information

© International Society for Plant Molecular Biology 1993

Authors and Affiliations

  • Olivier Panaud
    • 1
  • Gerard Magpantay
    • 2
  • Susan McCouch
    • 1
  1. 1.Plant Breeding and Biometry DepartmentCornell UniversityIthacaUSA
  2. 2.Plant Breeding Division, Genome Mapping LaboratoryInternational Rice Research InstituteManilaPhilippines

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