TaqmanMGB探针冻融稳定性研究  

Study on Freeze-thaw Cycles on Taqman MGB Stability

作　　者：臧超[1] 王晶[1] 李亮[1] 隋志伟[1] 余笑波[2] 黎朋[1] 

机构地区：[1]中国计量科学研究院,北京100013 [2]中国计量学院,浙江,杭州310018

出　　处：《计量学报》2011年第B12期96-100,共5页Acta Metrologica Sinica

基　　金：国家转基因生物新品种培育科技重大专项（2008ZX08012003）;国家科技支撑计划项目（2008BAK41801）.

摘　　要：TaqmanMGB探针的稳定性是实时荧光定量PCR准确定量的关键因素之一。然而在实验过程中，常常因为运输、实验室保存或使用条件的变化，造成探针的反复冻融，影响实验结果。采用高效液相色谱技术定量研究了反复冻融的MGB探针的稳定性；并比较分析了冻融探针对qPCR的影响程度。结果表明：MGB探针可以耐受30次以内的反复冻融，随着探针浓度的降低，探针反复冻融后qPCR量值波动区间增大，因此，为保证结果稳定性，qPCR实验使用的MGB探针应尽量减少反复冻融次数。The stability of Taqman MGB probes is crucial for the accuracy of qPCR quantification. However, the stability is easy to be affected by multi freeze thawing process of transportations, storage surroundings and usage conditions. The freeze thawing stability of MGB probe is quantified measured by HPLC and the influence to qPCR is evaluated with multi freeze thawing probes. The results show that a 30-time freeze thawing process is tolerable for MGB probes. QPCR quantitation accuracy decreases together with probe concentration. Therefore, reducing freeze thawing process of MGB probes is offered to keep the stability of MGB probes when experiments.

关 键 词：计量学 MGB探针 反复冻融 高效液相色谱 实时荧光定量PCR 

分 类 号：TB99[一般工业技术—计量学]