Rapid Nucleic Acid Extraction and Purification Using a Miniature Ultrasonic Technique

• Main Authors: Darren W. Branch, Erika C. Vreeland, Jamie L. McClain, Jaclyn K. Murton, Conrad D. James, Komandoor E. Achyuthan
• Format: Article
• Language: English
• Published: MDPI AG 2017-07-01
• Series: Micromachines
• Subjects: acoustic lysis; microfluidic; nucleic acids; extraction; purification; separation; Point-of-Care
• Online Access: https://www.mdpi.com/2072-666X/8/7/228

Miniature ultrasonic lysis for biological sample preparation is a promising technique for efficient and rapid extraction of nucleic acids and proteins from a wide variety of biological sources. Acoustic methods achieve rapid, unbiased, and efficacious disruption of cellular membranes while avoiding the use of harsh chemicals and enzymes, which interfere with detection assays. In this work, a miniature acoustic nucleic acid extraction system is presented. Using a miniature bulk acoustic wave (BAW) transducer array based on 36° Y-cut lithium niobate, acoustic waves were coupled into disposable laminate-based microfluidic cartridges. To verify the lysing effectiveness, the amount of liberated ATP and the cell viability were measured and compared to untreated samples. The relationship between input power, energy dose, flow-rate, and lysing efficiency were determined. DNA was purified on-chip using three approaches implemented in the cartridges: a silica-based sol-gel silica-bead filled microchannel, nucleic acid binding magnetic beads, and Nafion-coated electrodes. Using E. coli, the lysing dose defined as ATP released per joule was 2.2× greater, releasing 6.1× more ATP for the miniature BAW array compared to a bench-top acoustic lysis system. An electric field-based nucleic acid purification approach using Nafion films yielded an extraction efficiency of 69.2% in 10 min for 50 µL samples.