CAJ | 학술논문

目的：确定聚合酶链反应－限制性片段长度多态性（ Polymerase chain reaction-restricted fragment length polymor-phism，PCR-RFLP）技术检测二甲基精氨酸二甲胺水解酶2（dimethyl arginine dimethylamine hydrolase 2，DDAH2）基因－449G／C的最适实验条件。方法对影响DDAH2基因PCR反应的主要因素进行研究。结果最适变性温度为95℃，最适退火温度为58℃，最适引物浓度为0.25μmol／L，最适2X Power Taq PCR Master Mix浓度为12.5μl，酶切体系为15.5μl体系中加5.0μl产物用5 U的限制性内切酶Sma I消化。结论应用PCR-RFLP技术建立的DDAH2基因－449 G／C位点多态位点检测方法简便、经济、快速。
목적：학정취합매련반응－한제성편단장도다태성（ Polymerase chain reaction-restricted fragment length polymor-phism，PCR-RFLP）기술검측이갑기정안산이갑알수해매2（dimethyl arginine dimethylamine hydrolase 2，DDAH2）기인－449G／C적최괄실험조건。방법대영향DDAH2기인PCR반응적주요인소진행연구。결과최괄변성온도위95℃，최괄퇴화온도위58℃，최괄인물농도위0.25μmol／L，최괄2X Power Taq PCR Master Mix농도위12.5μl，매절체계위15.5μl체계중가5.0μl산물용5 U적한제성내절매Sma I소화。결론응용PCR-RFLP기술건립적DDAH2기인－449 G／C위점다태위점검측방법간편、경제、쾌속。
Objectives To determine the optimum experimental conditions for DDAH2 gene-449G/C polymorphism by Poly-merase chain reaction-restricted fragment length polymorphism ( PCR-RFLP).Methods The main influential factors were detected by PCR in electrophoresis, the main factors were detected by RFLP in different system.Results The optimum conditions:denatur-ation temperature was 95℃, annealing temperature was 58℃, Stretching temperature was 72℃, primer concentration was 0.25μmol/L, 2X Power Taq PCR Master Mix was 12.5μl, the effective system was 15.5μl including 5μl DNA solution and 5U en-zyme.Conclusions It suggested that the method of detecting the dimethyl arginine dimethylamine hydrolase 2 gene-449G/C poly-morphism based on PCR-RFLP theory is facilitated, economic, and rapid.