A bifurcated continuous field-flow fractionation (BCFFF) chip for high-yield and high-throughput nucleic acid extraction and purification. Issue 22 (17th October 2019)

• Record Type: Journal Article
• Authors: Zhang, Chenguang; Sun, Gongchen; Senapati, Satyajyoti; Chang, Hsueh-Chia
• Is Part Of: Lab on a chip. Volume 19:Issue 22(2019)
• Page Start: 3853
• Page End: 3861
• Publication Date: 2019-10-17
• Subjects: Miniature electronic equipment -- Periodicals; Combinatorial chemistry -- Periodicals; Biotechnology -- Periodicals; 543.0813
• DOI: 10.1039/c9lc00818g
• Languages: English
• ISSNs: 1473-0197

Abstract

Abstract : We report a new Bifurcated Continuous Field-Flow Fractionation (BCFFF) microfluidic chip for isolation and purification of nucleic acids from blood plasma with high and concentration-independent yield. The platform is ideal for isolation and quantification of small miRNAs. Abstract : We report a bifurcated continuous field-flow fractionation (BCFFF) chip for high-yield and high-throughput (20 min) extraction of nucleic acids from physiological samples. The design uses a membrane ionic transistor to sustain low-ionic strength in a localized region at a junction, such that the resulting high field can selectively isolate high-charge density nucleic acids from the main flow channel and insert them into a standardized buffer in a side channel that bifurcates from the junction. The high local electric field and the bifurcated field-flow design facilitate concentration reduction of both divalent cation (Ca 2+ ) and molecular PCR inhibitors by more than two orders of magnitude, even with high-throughput continuous loading. The unique design with a large (>20 mM mm −1 ) on-chip ionic-strength gradient allows miniaturization into a high-throughput field-flow fractionation chip that can be integrated with upstream lysing and downstream PCR/sensor modules for various nucleic acid detection/quantification applications. A concentration-independent 85% yield for extraction and an overall post-PCR yield exceeding 60% are demonstrated for a 111 bp dsDNA in 10 μL of human plasma, compared to no amplification with the raw sample. A net yield four times larger than a commercial extraction kit is demonstrated for miR-39 in human plasma.