Adsorption and isolation of nucleic acids on cellulose magnetic beads using a three-dimensional printed microfluidic chip

Abstract

While advances in genomics have enabled sensitive and highly parallel detection of nucleic acid targets, the isolation and extraction of the nucleic acids remain a critical bottleneck in the workflow. We present here a simple 3D printed microfluidic chip that allows for the vortex and centrifugation free extraction of nucleic acids. This novel microfluidic chip utilizes the presence of a water and oil interface to filter out the lysate contaminants. The pure nucleic acids, while bound on cellulose particles, are magnetically moved across the oil layer. We demonstrated efficient and rapid extraction of spiked Human Papillomavirus (HPV) 18 plasmids in specimen transport medium, in under 15 min. An overall extraction efficiency of 61% is observed across a range of HPV plasmid concentrations (5 × 10(1) to 5 × 10(6) copies/100 μl). The magnetic, interfacial, and viscous drag forces inside the microgeometries of the chip are modeled. We have also developed a kinetics model for the adsorption of nucleic acids on cellulose functionalized superparamagnetic beads. We also clarify here the role of carrier nucleic acids in the adsorption and isolation of nucleic acids. Based on the various mechanistic insights detailed here, customized microfluidic devices can be designed to meet the range of current and emerging point of care diagnostics needs.

AirJump: Using Interfaces to Instantly Perform Simultaneous Extractions

“…By “jumping” PMP-bound analyte through a liquid/air phase interface and narrow (∼1 mm) air gap, we achieve a purification efficiency higher than that observed with traditional “tube-based” washing, due in part to the lack of surface-associated contaminants (e.g., background residue on the side of a washing tube, contaminants trapped by PMPs repeatedly drawn against the sidewall of a tube during washing steps, Figure 1 B). In previous ESP configurations, PMPs were drawn along a surface 6 , 7 , 9 − 12 , 16 − 19 or magnetically immobilized on a moving surface. 20 , 21 In contrast, analytes are extracted directly from the top surface of the sample volume, mitigating potential mechanisms of loss via friction and/or adsorption.…”

3D printed microfluidic mixer for point-of-care diagnosis of anemia

“…In contrast, a consumer-grade 3D printer can produce highly versatile, durable microfluidic POC devices without the need of laboratory settings[13–16]. Application of 3D printing in microfluidic technology has shown tremendous potential in tissue engineering[17–19], sample preparation techniques[20], and cell processing[21–24]. Zhang et al introduced a 3D printed microfluidic chip which utilizes a water and oil interface to filter out the lysate contaminants for extraction of nucleic acids[20].…”

“…Application of 3D printing in microfluidic technology has shown tremendous potential in tissue engineering[17–19], sample preparation techniques[20], and cell processing[21–24]. Zhang et al introduced a 3D printed microfluidic chip which utilizes a water and oil interface to filter out the lysate contaminants for extraction of nucleic acids[20]. Chan et al developed movable 3D printed microfluidic chip components, such as torque-actuated pump and valve, rotary valve, and pushing valve, for assembling microfluidic POC devices[25].…”

3D printed auto-mixing chip enables rapid smartphone diagnosis of anemia

“…[13][14][15][16] Application of 3D printing in microfluidic technology has shown tremendous potential in tissue engineering, [17][18][19] sample preparation a) Author to whom correspondence should be addressed. Electronic mail: meih@ksu.edu techniques, 20 and cell processing. [21][22][23][24] Zhang et al introduced a 3D printed microfluidic chip which utilizes a water and oil interface to filter out the lysate contaminants for extraction of nucleic acids.…”

“…[21][22][23][24] Zhang et al introduced a 3D printed microfluidic chip which utilizes a water and oil interface to filter out the lysate contaminants for extraction of nucleic acids. 20 Chan et al developed movable 3D printed microfluidic chip components, such as torque-actuated pump and valve, rotary valve, and pushing valve, for assembling microfluidic POC devices. 25 The tedious, multiple 3D printed components need to be assembled and employed for protein quantification from artificial urine samples with a smartphone as an imaging platform.…”