Digital PCR on an integrated self-priming compartmentalization chip.
An integrated on-chip valve-free and power-free microfluidic digital PCR device is for the first time developed by making use of a novel self-priming compartmentalization and simple dehydration control to realize 'divide and conquer' for single DNA molecule detection. The high gas solubility of PDMS is exploited to provide the built-in power of self-priming so that the sample and oil are sequentially sucked into the device to realize sample self-compartmentalization based on surface tension. The lifespan of its self-priming capability was about two weeks tested using an air-tight packaging bottle sealed with a small amount of petroleum jelly, which is significant for a practical platform. The SPC chip contains 5120 independent 5 nL microchambers, allowing the samples to be compartmentalized completely. Using this platform, three different abundances of lung cancer related genes are detected to demonstrate the feasibility and flexibility of the microchip for amplifying a single nucleic acid molecule. For maximal accuracy, within less than 5% of the measurement deviation, the optimal number of positive chambers is between 400 and 1250 evaluated by the Poisson distribution, which means one panel can detect an average of 480 to 4804 template molecules. This device without world-to-chip connections eliminates the constraint of the complex pipeline control, and is an integrated on-chip platform, which would be a significant improvement to digital PCR automation and more user-friendly.
Research Center for Analytical Instrumentation, Institute of Cyber-Systems and Control, State Key Laboratory of Industrial Control Technology, Zhejiang University, Hangzhou, PR China. muying@zju.edu.cn.
Qiangyuan
Q
Zhu
Lin
L
Qiu
Bingwen
B
Yu
Yanan
Y
Xu
Yibo
Y
Gao
Tingting
T
Pan
Qingchang
Q
Tian
Qi
Q
Song
Wei
W
Jin
Qinhan
Q
Jin
Ying
Y
Mu
D045744:Cell Line, Tumor; D004273:DNA, Neoplasm / Q000737:chemistry*; D006801:Humans; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000295:instrumentation* / Q000379:methods*
D004273:DNA, Neoplasm