Welcome to Bulletin of Botanical Research! Today is Share:

Bulletin of Botanical Research ›› 2006, Vol. 26 ›› Issue (1): 93-97.doi: 10.7525/j.issn.1673-5102.2006.01.022

Previous Articles     Next Articles

Optimization of RAMP-PCR reaction system in genomic DNA of radish (Raphanus sativus)

GONG Yi-Qin;LI Pei;WANG Ming-Xia;ZHAO Li-Ping;LIU Li-Wang*   

  1. National Key Laboratory of Crop Genetics and Germplasm Enhancement; College of Horticulture, Nanjing Agricultural University, Nanjing 210095
  • Received:1900-01-01 Revised:1900-01-01 Online:2006-01-20 Published:2006-01-20
  • Contact: LIU Li-Wang
  • Supported by:

Abstract: Genomic DNA of Radish was analyzed by optimizing the concentration of Mg2+, dNTPs and primer in Random Amplified Microsatellite Polymorphism (RAMP) system. Three different concentration gradients were set respectively, Mg2+ 0.75,1.5,3.0 mmol·L-1;dNTPs 0.05,0.15,0.3 mmol·L-1;primer 0.065,0.2,0.4 μmol·L-1, and the suitable annealing temperature was also screened. An optimization of RAMP reaction system for radish genomic DNA is(20 μL): dNTPs 0.15 mmol·L-1,Mg2+ 1.5 mmol·L-1,primer 0.2~0.4 μmol·L-1,DNA 10 ng,Taq E 0.8U.The amplification protocol was: 94℃ 3 min,42 cycles for 1 min at 94℃,1 min at 45℃, 1.5 min at 72℃, followed by a final extension of 8 min at 72℃. The suitable system was applied in screening the primer combinations, and the genetic diversity of seven radish varieties was analyzed in RAMP marker system.

Key words: radish, RAMP marker, system optimization, genetic diversity

CLC Number: