摘要
基于固相萃取原理,设计并制作了一种集成核酸提取功能的微流控芯片,用于人体全血中脱氧核糖核酸(DNA)的提纯。芯片主要包括:混合微通道、裂解腔、DNA提取腔、DNA存储腔。采用3D打印技术制作出芯片模板,通过模板注塑成型、氧等离子体键合等工艺制作出微流控芯片。利用该芯片,可以在20 min内完成血液和试剂的顺序加载、快速混合、血细胞裂解、DNA的提取等操作,且试剂消耗量少。通过对所提DNA链上稳定表达的内参基因甘油醛-3-磷酸脱氢酶(GAPDH)进行PCR扩增,并对扩增产物进行熔解曲线分析,以验证所提取DNA的质量。
We designed and fabricated an integrated nucleic acid extraction microfluidic chip based on solid phase extraction（ SPE）,which is used for the extraction and purification of deoxyribonucleic acid（ DNA） from human whole blood. The chip mainly includes mixing micro-channel,lysis chamber,DNA extraction chamber,and storage chamber. The templates of the chip were fabricated by a 3D printer. The microfluidic chip was made by injection molding and bonded by oxygen plasma technique. The chip has been successfully used for sequential injection of blood and reagent,rapid mixing of blood and reagent,lysis of blood cell,DNA extraction,etc.These operations can be accomplished in 20 minutes with less consumption of reagents. The reference gene of the extracted DNA, Glyceraldehyde-3-phosphate dehydrogenase（ GAPDH）, is amplified by polymerase chain reaction（ PCR） successfully. The amplicons were analyzed by melting curves in order to verify the quality of the extracted DNA. The results indicate that the microfluidic chip can be used to extract and purify genomic DNA,with high quality,from human whole blood.
出处
《分析试验室》
CAS
CSCD
北大核心
2015年第10期1231-1234,共4页
Chinese Journal of Analysis Laboratory
基金
中科院战略性先导科技专项基金(XDA08040109)资助
关键词
3D打印
微流控芯片
核酸提取
顺序注射
3D printing
Microfluidic chip
Nucleic acid extraction
Sequential injection
