양파(Allium cepa L)의 RAPD 분석조건 최적화에 관한 연구

Optimization of RAPD-PCR Conditions for Onions, Allium cepa L.

  • 정순재 (동아대학교 생명자원과학부) ;
  • 양보경 (동아대학교 생명자원과학부) ;
  • 김익수 (동아대학교 생명자원과학부) ;
  • 박선희 (동아대학교 생명자원과학부) ;
  • 서전규 (경상남도 농업기술원 양파시험장) ;
  • 남재성 (동아대학교 생명자원과학부) ;
  • 김현경 (동아대학교 생명자원과학부) ;
  • 김도훈 (동아대학교 생명자원과학부)
  • 발행 : 2000.04.01

초록

The optimized RAPD-PCR conditions, which can be utilized as a basic information for the analysis of the genetic characteristics were investigated with four onion varieties, named Changryungdaego, Yeoeuijuhwang, Yakwangju, and Dabonghwang using Operon primers, OPR01 (TGCGGGTCCT) and OPZ20 (ACTTTGGCGG). We tested several concentrations of DNA, primer, and MgCl2, annealing temperature, number of PCR cycle, and presence/absence of pre-heating time at the begining of PCR reation in the 25${mu}ell$ volume. The best RAPD profiles were obtained using 50ng of DNA, 5mM of primer, 1.5mM of MgCl2, 45$^{\circ}C$ of annealing temperature and an absence of pre-heating time. An establishment of the stable and reproducible RAPD-PCR conditions are expected to be useful for the subsequent RAPD-related investigation, such as genetic characterization of the onion strains, re-establishment of phylogenetic relationships and development of new varieties.

키워드

참고문헌

  1. Korean. J. Breed v.29 no.3 Analysis of genetic Relationships among Glycine species using RAPDs Baek, I. Y.;Y. H. Yoon;D. C. Shin;G. H. Park;Y. H. Hwang;D. U. Kim
  2. BioTechnique v.18 no.5 Reproducible amplification of RAPA markers from Vertebrate DNA Bielawski, J. P.;K. Noack;D. E. Pumo
  3. Korean J. Breed v.27 no.3 Protein and Lipid contents of Kangwon local soybean and their RAPDs Heo, N. K.;D. H. Park;K. M. Shim;S. Y. Lee;K. S. Kim;N. S. Kim
  4. Theor. Appl. Genet. v.90 Random amplified polymorphic DNA variation in the eggplant Salanum Melongena L.(Solanaceae) Karibaloo, J. L.;S. Brauner;L. D. Gottlieb
  5. Agri. Inst. Cooperation v.37 Determination of the genetic purity of F1 Hybrid seed in Watermelon(citrullus pulgaris)by using random amplified polymorphic DNA Kim, B. D.;B. C. Kang;J. Y. Kim
  6. Korean J. Breed v.30 no.3 RARD analysis in Korean native and introduced Chinese Chives(Allium spp.) Kwon, Y. C.;W. Y. Han;B. J. Kim;C. S. Jung;Y. H. Kwak
  7. Theor. Appl. Genet v.85 Identification of apple cultivars using RAPD markers Koller, B. A;J. M. Lehmann;C. Gessler
  8. Kor. J. Hort. Sci. & Tech v.16 no.1 RAPD analysis for genetic diversity of Melon species Mo, S. Y.;S. H. Im;G. D. Go;C. M. Chong;D. H. Kim
  9. PCR methods and application v.1 The random amplified polymorphic DNA for fingerprinting Plants Munthali, M.;B. V. Ford-Lioyd;H. J. Newbury
  10. Kor. Hort. Abst. v.16 no.4 Classification of Capsium annumm germplasm using random amplified polymorphic DNA Nam, S. H.;G. W. Choi;I. W. Yoo
  11. Korean J. Biol. Sci. v.3 Identification of a tandemly repeated DNA sequence using combined RAPD and FISH in Welsh Onion (Allium Fistulosum) Seo, B. B.;G. S. Do;S. H. Lee
  12. Korean J. Breed v.27 no.1 Genetic diversity in Watermelon(Cirullus vulgaris L.)germplasm through RAPA analysis Shin, J. S.;S. J. Lee;K. W. Park
  13. Theor. Appl. Genet. v.85 Using randomly amplified polymorphic DNA for evaluating genetic relationships among papaya cultivars Stiles, J. I.;C. Lemme;S. Sondur;M. B. Morshidi;R. Manshardt
  14. Joint Plant Breeding Symposia Series Identification and reability of RAPD marker Weeden, N. F.;G. M. Timmerman;M. Hemmat;B. F. Kneen;M. A. Lodhi
  15. Theor. Appl. genet. v.86 Random amplified polymorphic DNA(RAPD) marker for genetic analysis in Allium Wilkie, S. E.;P. G. Issac;R. J. Slater
  16. Nucl. Acids Res. v.18 DNA polymorphisms amplified by arbitrary primers are useful as genetic markers Wiiliams, J. G. K.;A. R. Kubelik;K. J. Livak;J. A. Rafalski;S. V. Tingey
  17. J. Kor. Soc. Hort. Sci. v.36 no.5 Factor influencing RAPD band patterns in Malus domestica Yae, B. W.;H. Y. Park;Y. U. Shin;D. K. Lee;J. H. Kim;K. C. Ko
  18. Korean J. Breed v.30 no.2 Optimization of the random amplified polymorphic DNA analyses procedure in Capsicum annuum L. Yang, T. J.;H. G. Park.