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Title: Rapid Nucleic Acid Extraction and Purification Using a Miniature Ultrasonic Technique

Abstract

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 50more » µL samples.« less

Authors:
 [1];  [2];  [3];  [4];  [5];  [1]
  1. Sandia National Lab. (SNL-NM), Albuquerque, NM (United States). Nano and Micro Sensors Dept.
  2. Imagion Biosystems, Inc., Albuquerque, NM (United States)
  3. Sandia National Lab. (SNL-NM), Albuquerque, NM (United States). MEMS Technologies Dept.
  4. Sandia National Lab. (SNL-NM), Albuquerque, NM (United States). Bioenergy and Defense Technologies Dept.
  5. Sandia National Lab. (SNL-NM), Albuquerque, NM (United States). Physics Based Microsystems Dept.
Publication Date:
Research Org.:
Sandia National Lab. (SNL-NM), Albuquerque, NM (United States)
Sponsoring Org.:
USDOE National Nuclear Security Administration (NNSA)
OSTI Identifier:
1399498
Report Number(s):
SAND-2017-3875J
Journal ID: ISSN 2072-666X; 652447
Grant/Contract Number:  
NA0003525
Resource Type:
Accepted Manuscript
Journal Name:
Micromachines
Additional Journal Information:
Journal Volume: 8; Journal Issue: 7; Journal ID: ISSN 2072-666X
Publisher:
MDPI
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 77 NANOSCIENCE AND NANOTECHNOLOGY; acoustic lysis; microfluidic; nucleic acids; extraction; purification; separation; Point-of-Care

Citation Formats

Branch, Darren W., Vreeland, Erika C., McClain, Jamie L., Murton, Jaclyn K., James, Conrad D., and Achyuthan, Komandoor E. Rapid Nucleic Acid Extraction and Purification Using a Miniature Ultrasonic Technique. United States: N. p., 2017. Web. doi:10.3390/mi8070228.
Branch, Darren W., Vreeland, Erika C., McClain, Jamie L., Murton, Jaclyn K., James, Conrad D., & Achyuthan, Komandoor E. Rapid Nucleic Acid Extraction and Purification Using a Miniature Ultrasonic Technique. United States. doi:10.3390/mi8070228.
Branch, Darren W., Vreeland, Erika C., McClain, Jamie L., Murton, Jaclyn K., James, Conrad D., and Achyuthan, Komandoor E. Fri . "Rapid Nucleic Acid Extraction and Purification Using a Miniature Ultrasonic Technique". United States. doi:10.3390/mi8070228. https://www.osti.gov/servlets/purl/1399498.
@article{osti_1399498,
title = {Rapid Nucleic Acid Extraction and Purification Using a Miniature Ultrasonic Technique},
author = {Branch, Darren W. and Vreeland, Erika C. and McClain, Jamie L. and Murton, Jaclyn K. and James, Conrad D. and Achyuthan, Komandoor E.},
abstractNote = {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.},
doi = {10.3390/mi8070228},
journal = {Micromachines},
number = 7,
volume = 8,
place = {United States},
year = {2017},
month = {7}
}

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