摘要
微流控荧光PCR,产物在高分辨率熔解(HRM)曲线分析的过程中,随着DNA的解链荧光强度发生突变,而能否检测到该突变点则依赖于温控的精度。提出了多阈值PID算法,设计了一种适用于高分辨率熔解曲线分析的FPGA温度控制系统。根据测得温度值与设定值之间的差异,改变FPGA电路系统输出的脉冲宽度调制(pulse width modulation,PWM)波占空比和算法的调节频率实现对温度的准确控制。经过温度标定后,系统温度控制精度优于0.1℃,分辨率达到±0.01℃。HRM分析实验结果表明,本方法成功地辨别出了DNA序列的单碱基差异,温度控制系统达到了设计的要求。
In the analysis of high resolution melting （HRM） of microfluidic fluorescence PCR amplification products, fluorescence intensity has a sudden change with the delink of DNA chain, while whether the abrupt change can be detected is dependent on the accuracy of temperature control. A kind of multi-threshold PID algorithm was put forward, and a temperature control system based on FPGA was designed to adapt to the analysis of high resolution melting. Based on the difference of the measurement value and the temperature change can be controlled accurately through changing the modulation （PWM） wave and the control frequency of the algorithm from t pulse-time ratio he FPGA circuit the set value, of pulse width system. After temperature calibrations of the measurement system, the accuracy precedes 0.1 ℃ and the resolution achieves ±0.01 ℃. The result of HRM experiment shows that this system can detect the single base mutation of DNA sequence successfully, and the temperature control system can meet the design requirements
出处
《大气与环境光学学报》
CAS
CSCD
2015年第5期417-424,共8页
Journal of Atmospheric and Environmental Optics
基金
中国科学院战略性先导科技专项基金项目(XDA08040109)资助
关键词
微流控
铂电阻
多阈值PID算法
脉冲宽度调制波
高分辨率熔解
microfluidic
platinum resistance
multi-threshold PID algorithm
pulse width modulation wave
high resolution melting
