论文部分内容阅读
以DNA为模板合成的荧光银纳米簇(DNA-Ag NC)是一类性能优秀的发光探针,已被广泛应用于传感与成像领域.目前大多数定量方法都是基于目标物结合带来的直接的银纳米簇发光强度的变化,因此,分析方法的重现性将不可避免地受实验场地或测定条件变化的影响.本工作中,采用以G-四链体为模板的银纳米簇(GQ-AgNC)与菁染料Cyanine5(Cy5)为能量供-受体对,发展了基于荧光共振能量转移(FRET)机理的mi RNA比率测定方法.其中,Cy5标签具有通用性,简化了实际应用中的实验设计,降低了试剂成本.针对mi RNA let-7a的方法,检测动态范围为12~300nmol/L,检测限为6.9 nmol/L,且可以将let-7a与let-7家族中的其他mi RNA区分.HepG2细胞的总RNA提取物加标回收结果表明该方法具有应用于临床样本测试的潜力.本研究工作拓展了DNA-Ag NC的应用,并有助于加深在FRET设计中以DNA-Ag NC为供体进行分析应用的进一步理解.
Fluorescent silver nanoclusters (DNA-Ag NCs) synthesized with DNA are a class of excellent luminescent probes and have been widely used in the field of sensing and imaging.At present, most quantitative methods are based on the combination of target Of the direct silver nanocluster luminescence intensity changes, therefore, the reproducibility of analytical methods will inevitably be affected by changes in the experimental site or measuring conditions.In this work, the use of G-quadruplex as a template of silver nanoclusters (GQ-AgNC) with cyanine Cyanine5 (Cy5) as energy donor-acceptor pairs, a miRNA ratio assay based on the fluorescence resonance energy transfer (FRET) mechanism has been developed, in which the Cy5 label is versatile and simplifies practical applications In the experimental design, the cost of the reagent was reduced.The detection limit of mi RNA let-7a was 12 ~ 300 nmol / L, the detection limit was 6.9 nmol / L, and let-7a and let- Other mi RNA discrimination.The spike recovery results of total RNA extracts from HepG2 cells indicate that this method has the potential to be used in clinical sample testing.The work of this work broadens the application of DNA-Ag NCs and helps to deepen the understanding of DNA-Ag NC for the donor analysis and application of further management solution.