Application Research Of Microscale Nucleic Acid Based On Microfluidic Chips Technology

Microfluidic Chip is a precise experiment technology which can control the microfluidics in micro channel. It integrates many reaction steps invoved in the chemistry and biology fields highly on a scale of a few square centimeters or even a few square microns. This technology is widely used in genomic sequencing, diseases, diagnosis and therapy, drug sorting and screening, and other fields, because it is characteristic of precise control samples, trace the reagent consumption, good biocompatibility, high reaction efficiency, high throughput analysis, lower production cost, highly integrated automation etc. It is one of the most important advanced analysis technologies in the 21 st century.With the beginning of the Precision Medicine Initiative, human's exploration of the tumor is not only confined to study the apparent characteristics of tumor. Based on tumor genome, transcriptome, and proteome of trace sample for further studies will help the researchers fundamentally found tumor of generation, evolutionary process and specific target of tumor. Thus, how to make these trace samples be tested more accurate, more efficient and cheaper is the first problem in the field of cancer research.Based on the design and processing of the Microfluidic Chip technology, cast molding manufacture successfully Microdroplets Microfluidic Chip which is made from PDMS, and set up a standardized Microfluidic Chip system. Through a microdroplets generation experiments, we can find the main factors which influenced by microdroplets generation are continuous phase and disperse phase under the situation of microchannel structure and coefficient of viscosity ratio, and determine the curve fit expression of the microdroplets diameter. Meanwhile, applying this system, it can manufacture p L~n L(0.81 p L~0.26 n L) microdroplets and can generate 103~106 droplets per minute.Through applying this system, we do experiments for microscale nucleic acid q PCR amplification experiments. Under the comparison between this system and traditional q PCR experiment, we can find this system has many advantages, such as high amplification efficiency(Ct value appear in advance 2 ~ 7 cycles), low amplification limit(10-8~10-3 ng/?L concentration of DNA template). It provides a solution which is based on nucleic acid PCR amplification. What's more, it provide a theoretical basis for Single-cell whole genome amplification, library construction of metagenome and other molecular diversity advanced technology.