CAJ | 학술논문

　　为了探讨广西甜茶ISSR实验中的多种因素对实验结果的影响，采用单因素法对PCR反应体系中的5个潜在因素(Mg2+, dNTP,引物, Taq酶和模板DNA)在5水平上进行优化实验，并进一步优化了反应程序中的退火温度和循环次数。结果表明：25滋L的最佳反应体系为，1×PCR Buffer、MgCl22.0 mmol/L、dNTP 0.2 mmol/L、引物0.8滋mol/L、Taq DNA聚合酶0.75 U、模板DNA 80 ng；最佳反应程序为：在94℃下进行3 min预变性；随后循环扩增35次(包括94℃变性1 min,54℃退火1 min,72℃延伸1 min)；最后在72℃下延伸10 min。本研究建立的广西甜茶的最佳ISSR-PCR反应条件为进一步进行遗传多样性研究奠定了基础。
　　위료탐토엄서첨다ISSR실험중적다충인소대실험결과적영향，채용단인소법대PCR반응체계중적5개잠재인소(Mg2+, dNTP,인물, Taq매화모판DNA)재5수평상진행우화실험，병진일보우화료반응정서중적퇴화온도화순배차수。결과표명：25자L적최가반응체계위，1×PCR Buffer、MgCl22.0 mmol/L、dNTP 0.2 mmol/L、인물0.8자mol/L、Taq DNA취합매0.75 U、모판DNA 80 ng；최가반응정서위：재94℃하진행3 min예변성；수후순배확증35차(포괄94℃변성1 min,54℃퇴화1 min,72℃연신1 min)；최후재72℃하연신10 min。본연구건립적엄서첨다적최가ISSR-PCR반응조건위진일보진행유전다양성연구전정료기출。
In order to study the influence of several potential factors on ISSR system for Rubus suavissimus S. Lee., five factors (Mg2+, dNTP, primer, Taq DNA polymerase and DNA template) were optimized by multiple single factor experiments at 5 concentration levels, annealing temperatures and amplification cycle numbers were also optimized. The results showed that the optimum 25μL reaction system comprises 1íPCR buffer, MgCl2 2.0 mmol/L, dNTP 0.2 mmol/L, primer 0.8μmol/L, Taq DNA polymerase 0.75 U and template DNA 80 ng;and the optimum PCR procedure is as an initial pre-denaturing at 94℃for 3 min, which precedes 35 cycles of denaturing at 94℃for 1 min, annealing at 54℃ for 1 min, and extension at 72℃ for 1 min, with a final extension at 72℃ for 10 min. Establish-ment of the optimum ISSR-PCR systems and conditions would serve as a base for further study on the genetic diver-sity of Rubus suavissimus S. Lee.