CAJ | 학술논문

采用 L16(45)正交实验设计,对杉木 SRAP-PCR 反应体系中 Mg2+、dNTPs、Taq DNA 聚合酶用量、引物浓度及模板 DNA 用量等5个因素进行了优化,并确立了适宜杉木的最佳 SRAP-PCR 反应体系。杉木的 SRAP-PCR最佳反应体系为：反应体系总体积为25μL,含2.0 mmol/L Mg2+、0.3 mmol/L dNTPs 、0.3μmol/L 引物,1.0 U Taq DNA 聚合酶、50 ng 模板 DNA 及1×PCR buffer。各因素对杉木基因组 DNA SRAP-PCR 扩增结果的影响程度不同,其中 dNTPs 浓度影响最大,Mg2+浓度的影响最小。运用杉木单株基因组 DNA 对优化 SRAP-PCR 反应体系进行验证,均获得多态性丰富、条带清晰的扩增图谱,表明所确立的杉木 SRAP-PCR 反应体系稳定可靠。
채용 L16(45)정교실험설계,대삼목 SRAP-PCR 반응체계중 Mg2+、dNTPs、Taq DNA 취합매용량、인물농도급모판 DNA 용량등5개인소진행료우화,병학립료괄의삼목적최가 SRAP-PCR 반응체계。삼목적 SRAP-PCR최가반응체계위：반응체계총체적위25μL,함2.0 mmol/L Mg2+、0.3 mmol/L dNTPs 、0.3μmol/L 인물,1.0 U Taq DNA 취합매、50 ng 모판 DNA 급1×PCR buffer。각인소대삼목기인조 DNA SRAP-PCR 확증결과적영향정도불동,기중 dNTPs 농도영향최대,Mg2+농도적영향최소。운용삼목단주기인조 DNA 대우화 SRAP-PCR 반응체계진행험증,균획득다태성봉부、조대청석적확증도보,표명소학립적삼목 SRAP-PCR 반응체계은정가고。
The SRAP-PCR reaction system of Cunninghamia lanceolata was established by the orthogonal experiment design L1 6 (4 5 ).Five factors including Mg2 + concentration,dNTPs concentration,primer concentration,Taq DNA polymerase amount and template DNA amount of the SRAP-PCR reaction sys-tem were optimized,and the optimal SRAP-PCR reaction system suitable for genomic DNA of Cunning-hamia lanceolata was also established.The optimal SRAP-PCR reaction system was as follows:total vol-ume 25 μL,containing 2.0 mmol/L Mg2 + ,0.3 mmol/L dNTPs,0.3 μmol/L primer,50 ng template DNA,1.0 U Taq DNA polymerase and 1 x PCR buffer.The effects of five factors on amplification result of SRAP-PCR were different,in which the effect of dNTPs concentration was the greatest,but the effect of Mg2 + concentration was the least.The optimal SRAP-PCR reaction system was verified by means of genomic DNA of Cunninghamia lanceolata ,and the amplification pattern with rich polymorphism and clear band was obtained.It could be concluded that this SRAP-PCR reaction system used for genomic DNA of Cunninghamia lanceolata was steady and reliable.