CAJ | 학술논문

为了筛选适宜金龟甲RAPD-PCR的基因组DNA提取方法，分别采用改良的SDS法、平衡酚法、裂解液法和CTAB法提取棕色鳃金龟(HolotrichiatitanisReitter)的基因组DNA，通过DNA沉淀性状观察、产量和质量分析，以及随机引物PCR (RAPD-PCR)比较筛选最优的金龟甲基因组DNA提取方法。研究结果表明，改良的SDS法提取的基因组DNA产量较高，但纯度较低，并且降解严重，RAPD-PCR扩增谱带弥散较严重；裂解液法提取的基因组DNA纯度较高，降解程度较低，但产量最低，RAPD-PCR扩增谱带弥散较严重，并且存在非特异扩增现象；CTAB法提取的基因组DNA不仅产量和纯度较低，降解严重，而且RAPD-PCR扩增结果不稳定，具非特异扩增现象；平衡酚法提取的基因组DNA产量和纯度高，降解轻微，RAPD-PCR扩增结果稳定，扩增谱带弥散轻微。因此，在上述4种方法中，平衡酚法是最适合PAPD-PCR的金龟甲基因组DNA提取方法，裂解液法和改良的SDS法次之，CTAB法最差。
위료사선괄의금구갑RAPD-PCR적기인조DNA제취방법，분별채용개량적SDS법、평형분법、렬해액법화CTAB법제취종색새금구(HolotrichiatitanisReitter)적기인조DNA，통과DNA침정성상관찰、산량화질량분석，이급수궤인물PCR (RAPD-PCR)비교사선최우적금구갑기인조DNA제취방법。연구결과표명，개량적SDS법제취적기인조DNA산량교고，단순도교저，병차강해엄중，RAPD-PCR확증보대미산교엄중；렬해액법제취적기인조DNA순도교고，강해정도교저，단산량최저，RAPD-PCR확증보대미산교엄중，병차존재비특이확증현상；CTAB법제취적기인조DNA불부산량화순도교저，강해엄중，이차RAPD-PCR확증결과불은정，구비특이확증현상；평형분법제취적기인조DNA산량화순도고，강해경미，RAPD-PCR확증결과은정，확증보대미산경미。인차，재상술4충방법중，평형분법시최괄합PAPD-PCR적금구갑기인조DNA제취방법，렬해액법화개량적SDS법차지，CTAB법최차。
In order to select the proper extraction method of genome DNA suitable for RAPD-PCR from scarab, genome DNA of Holotrichia titanis Reitter was extracted by Improved SDS, Balance Phenol, Lysis Solution and CTAB Method, and the most proper extraction method was selected by observation on the character of DNA deposition, production & quality analyzing and RAPD-PCR. The results showed that the genome DNA extracted by Improved SDS Method was higher in production, but lower in purity, degraded seriously, and could generate seriously dispersed RAPD-PCR fingerprinting when amplifying with it as template. Genome DNA extracted by Lysis Solution Method was higher in purity, degraded slightly, but lowest in production, and could generate seriously dispersed and nontypical RAPD-PCR fingerprinting when amplifying with it as template. Genome DNA extracted by CTAB Method was not only lower in purity and production, degraded seriously, but also could not generate stable RAPD-PCR fingerprinting when amplifying with it as template, and there was nontypical amplification too. Genome DNA extracted by Balance Phenol Method was higher in purity and production, degraded slightly, and could generate stable RAPD-PCR fingerprinting without dispersion when amplifying with it as template. So among above four methods, Balance Phenol Method was the most suitable method to extract genome DNA from scarab, which was to be used for PAPD-PCR, Lysis Solution and Improved SDS Method took the second place, and CTAB Method was the most ill-fitted one.