World-to-digital-microfluidic interface enabling extraction and purification of RNA from human whole blood.
Digital microfluidics (DMF) is a powerful technique for simple and precise manipulation of microscale droplets of fluid. This technique enables processing and analysis of a wide variety of samples and reagents and has proven useful in a broad range of chemical, biological, and medical applications. Handling of "real-world" samples has been a challenge, however, because typically their volumes are greater than those easily accommodated by DMF devices and contain analytes of interest at low concentration. To address this challenge, we have developed a novel "world-to-DMF" interface in which an integrated companion module drives the large-volume sample through a 10 μL droplet region on the DMF device, enabling magnet-mediated recovery of bead-bound analytes onto the device as they pass through the region. To demonstrate its utility, we use this system for extraction of RNA from human whole blood lysates (110-380 μL) and further purification in microscale volumes (5-15 μL) on the DMF device itself. Processing by the system was >2-fold faster and consumed 12-fold less reagents, yet produced RNA yields and quality fully comparable to conventional preparations and supporting qRT-PCR and RNA-Seq analyses. The world-to-DMF system is designed for flexibility in accommodating different sample types and volumes, as well as for facile integration of additional modules to enable execution of more complex protocols for sample processing and analysis. As the first technology of its kind, this innovation represents an important step forward for DMF, further enhancing its utility for a wide range of applications.
Departments of Biotechnology and Bioengineering, ‡Systems Biology, and §Advanced Systems Engineering and Deployment, Sandia National Laboratories , Livermore, California, United States.
Mais J
MJ
Jebrail
Anupama
A
Sinha
Samantha
S
Vellucci
Ronald F
RF
Renzi
Cesar
C
Ambriz
Carmen
C
Gondhalekar
Joseph S
JS
Schoeniger
Kamlesh D
KD
Patel
Steven S
SS
Branda
D004867:Equipment Design; D006801:Humans; D007202:Indicators and Reagents; D046210:Microfluidic Analytical Techniques / Q000379:methods*; D044085:Microfluidics / Q000379:methods*; D012313:RNA / Q000097:blood* / Q000302:isolation & purification; D015203:Reproducibility of Results
D007202:Indicators and Reagents; D012313:RNA