CAJ | 학술논문

该研究分析普通PCR的不足，通过较多实验研究初步建立乳液PCR方法，改善PCR实验技术．采用人工合成的乳酸杆菌质粒为模板，不同成分配比条件下的乳液PCR和普通PCR对质粒序列进行扩增，对扩增产物进行琼脂糖凝胶电泳，电泳结束后对凝胶进行Gelred染色并用GS-800灰度扫描仪成像．进一步分析乳液PCR和普通PCR扩增产物的特异性以及不同成分配比条件下乳液PCR的扩增质量．通过比较两种PCR方法的扩增结果可得，在相同条件下，乳液PCR扩增特异性高于普通PCR扩增，并确定当水相中加入100g／LBSA，水油体积比在1：2．5时，破乳水饱和乙醚体积比为1：1时，水油相在1700rpm条件下连续混合5min时扩增效果最好．
해연구분석보통PCR적불족，통과교다실험연구초보건립유액PCR방법，개선PCR실험기술．채용인공합성적유산간균질립위모판，불동성분배비조건하적유액PCR화보통PCR대질립서렬진행확증，대확증산물진행경지당응효전영，전영결속후대응효진행Gelred염색병용GS-800회도소묘의성상．진일보분석유액PCR화보통PCR확증산물적특이성이급불동성분배비조건하유액PCR적확증질량．통과비교량충PCR방법적확증결과가득，재상동조건하，유액PCR확증특이성고우보통PCR확증，병학정당수상중가입100g／LBSA，수유체적비재1：2．5시，파유수포화을미체적비위1：1시，수유상재1700rpm조건하련속혼합5min시확증효과최호．
In the present study, the shortage of Amplification Complex Gene Sequence by Conventional PCR was analyzed, and then a protocol was constructed to minimize these problems. With the plasmid of Lactobacillus as a template, different components of the emulsion PCR and the conventional PCR plasmid template were effectively amplified to promote the amplified product with PCR to carry on agarose electrophoresis. With that accomplished, the Gelred dyeing was done to the gelatin and the image formation was obtained by a GS-800 gradation scanner. Then a further analysis was made to get the specialities of the emulsion PCR and the conventional PCR amplified products and the producing quality by emulsion PCR. In terms of different components, it was found out by contrasting the two methods that the speciality of the emulsion PCR amplification was higher than that of the conventional PCR with the same conditions. Moreover, when 100 g/1 BSA was added to aqueous phase with the volume ratio of water-in-oil(w/o) at 1:2.5, the volume ratio of breaking water-saturated diethyl ether at 1:1 and water and oil mixture was thoroughly mixed in 5 min at 1700 rpm, the best amplification effect was obtained.