CAJ | 학술논문

利用单因素梯度试验与正交试验相结合、方差分析与多重比较分析相结合的方法，对水仙简单重复序列-聚合酶链式反应（ SSR-PCR）扩增反应体系中的4个因素（ DNA、dNTPs、引物及Taq酶）进行优化分析，方差分析表明4个因素对SSR-PCR的影响均达到极显著水平，通过多重比较分析发现SSR引物和Taq酶对总体系的扩增结果影响最大。根据单因素梯度试验与正交试验结果，建立了适合水仙SSR标记分析的最优PCR反应体系：DNA模板2．5 ng·μL-1，10×PCR Buffer （含1．5 mmol· L-1 Mg2＋），dNTPs 0．25 mmol· L-1，上下游引物各0．50μmol· L-1，Taq酶1．00 U，总体积20μL。该优化体系的建立可为今后水仙SSR分析提供标准化的试验体系，为水仙种质鉴定、品种指纹图谱绘制、遗传多样性分析、群居遗传结构研究和遗传连锁图谱分析等领域提供技术支持。
이용단인소제도시험여정교시험상결합、방차분석여다중비교분석상결합적방법，대수선간단중복서렬-취합매련식반응（ SSR-PCR）확증반응체계중적4개인소（ DNA、dNTPs、인물급Taq매）진행우화분석，방차분석표명4개인소대SSR-PCR적영향균체도겁현저수평，통과다중비교분석발현SSR인물화Taq매대총체계적확증결과영향최대。근거단인소제도시험여정교시험결과，건립료괄합수선SSR표기분석적최우PCR반응체계：DNA모판2．5 ng·μL-1，10×PCR Buffer （함1．5 mmol· L-1 Mg2＋），dNTPs 0．25 mmol· L-1，상하유인물각0．50μmol· L-1，Taq매1．00 U，총체적20μL。해우화체계적건립가위금후수선SSR분석제공표준화적시험체계，위수선충질감정、품충지문도보회제、유전다양성분석、군거유전결구연구화유전련쇄도보분석등영역제공기술지지。
Four factors including DNA , dNTPs, primers and Taq-polymerase in simple sequence repeat-polymerase chain reaction ( SSR-PCR) of Narcissus were optimized .The comparison of single factor gradient experiments and orthogonal experimental design as well as analysis of variance and multiple comparison analysis were used in optimization .Analysis of variances results showed that the factors influenced SSR-PCR significantly .It was found that SSR primers and Taq-polymerase were the major factors by multiple com-parison analysis .According to the results of single factor gradient experiments and orthogonal experimental design , the optimization PCR system was established.It contained 2.5 ng·μL-1 DNA, 10 ×PCR Buffer (including 1.5 mmol· L-1 Mg2+), 0.25 mmol· L-1 dNTPs, 0.50 μmol· L-1 forward and reverse primers, and 1.00 U Taq-polymerase.This PCR system may offer a standardized experiment to analysis of SSR , and technical support to identification of variety , varieties of fingerprint mapping , analysis of genetic diversity, and genetic structure of social and genetic linkage maps study of Narcissus.