Literature DB >> 23061629

Genomic DNA extraction from cells by electroporation on an integrated microfluidic platform.

Tao Geng1, Ning Bao, Nammalwar Sriranganathanw, Liwu Li, Chang Lu.   

Abstract

The vast majority of genetic analysis of cells involves chemical lysis for release of DNA molecules. However, chemical reagents required in the lysis interfere with downstream molecular biology and often require removal after the step. Electrical lysis based on irreversible electroporation is a promising technique to prepare samples for genetic analysis due to its purely physical nature, fast speed, and simple operation. However, there has been no experimental confirmation on whether electrical lysis extracts genomic DNA from cells in a reproducible and efficient fashion in comparison to chemical lysis, especially for eukaryotic cells that have most of the DNA enclosed in the nucleus. In this work, we construct an integrated microfluidic chip that physically traps a low number of cells, lyses the cells using electrical pulses rapidly, then purifies and concentrates genomic DNA. We demonstrate that electrical lysis offers high efficiency for DNA extraction from both eukaryotic cells (up to ∼36% for Chinese hamster ovary cells) and bacterial cells (up to ∼45% for Salmonella typhimurium) that is comparable to the widely used chemical lysis. The DNA extraction efficiency has dependence on both the electric parameters and relative amount of beads used for DNA adsorption. We envision that electroporation-based DNA extraction will find use in ultrasensitive assays that benefit from minimal dilution and simple procedures.

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Year:  2012        PMID: 23061629      PMCID: PMC3492889          DOI: 10.1021/ac3026064

Source DB:  PubMed          Journal:  Anal Chem        ISSN: 0003-2700            Impact factor:   6.986


  60 in total

1.  High-throughput and real-time study of single cell electroporation using microfluidics: effects of medium osmolarity.

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Authors:  Paul Yager; Thayne Edwards; Elain Fu; Kristen Helton; Kjell Nelson; Milton R Tam; Bernhard H Weigl
Journal:  Nature       Date:  2006-07-27       Impact factor: 49.962

3.  Electroporation of mammalian cells in a microfluidic channel with geometric variation.

Authors:  Hsiang-Yu Wang; Chang Lu
Journal:  Anal Chem       Date:  2006-07-15       Impact factor: 6.986

4.  Microfluidic digital PCR enables multigene analysis of individual environmental bacteria.

Authors:  Elizabeth A Ottesen; Jong Wook Hong; Stephen R Quake; Jared R Leadbetter
Journal:  Science       Date:  2006-12-01       Impact factor: 47.728

5.  Using polarization-shaped optical vortex traps for single-cell nanosurgery.

Authors:  Gavin D M Jeffries; J Scott Edgar; Yiqiong Zhao; J Patrick Shelby; Christine Fong; Daniel T Chiu
Journal:  Nano Lett       Date:  2007-02       Impact factor: 11.189

6.  A pocket-sized convective PCR thermocycler.

Authors:  Nitin Agrawal; Yassin A Hassan; Victor M Ugaz
Journal:  Angew Chem Int Ed Engl       Date:  2007       Impact factor: 15.336

7.  Chitosan as a polymer for pH-induced DNA capture in a totally aqueous system.

Authors:  Weidong Cao; Christopher J Easley; Jerome P Ferrance; James P Landers
Journal:  Anal Chem       Date:  2006-10-15       Impact factor: 6.986

8.  A fully integrated microfluidic genetic analysis system with sample-in-answer-out capability.

Authors:  Christopher J Easley; James M Karlinsey; Joan M Bienvenue; Lindsay A Legendre; Michael G Roper; Sanford H Feldman; Molly A Hughes; Erik L Hewlett; Tod J Merkel; Jerome P Ferrance; James P Landers
Journal:  Proc Natl Acad Sci U S A       Date:  2006-12-11       Impact factor: 11.205

9.  A microfluidic flow-through device for high throughput electrical lysis of bacterial cells based on continuous dc voltage.

Authors:  Hsiang-Yu Wang; Arun K Bhunia; Chang Lu
Journal:  Biosens Bioelectron       Date:  2006-03-10       Impact factor: 10.618

Review 10.  Miniaturized PCR chips for nucleic acid amplification and analysis: latest advances and future trends.

Authors:  Chunsun Zhang; Da Xing
Journal:  Nucleic Acids Res       Date:  2007-06-18       Impact factor: 16.971

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  16 in total

1.  A mechanical cell disruption microfluidic platform based on an on-chip micropump.

Authors:  Yinuo Cheng; Yue Wang; Zhiyuan Wang; Liang Huang; Mingzhao Bi; Wenxiao Xu; Wenhui Wang; Xiongying Ye
Journal:  Biomicrofluidics       Date:  2017-04-04       Impact factor: 2.800

2.  Microfluidic approaches for cell-based molecular diagnosis.

Authors:  Dong Jun Lee; John Mai; Tony Jun Huang
Journal:  Biomicrofluidics       Date:  2018-09-14       Impact factor: 2.800

3.  Individually addressable multi-chamber electroporation platform with dielectrophoresis and alternating-current-electro-osmosis assisted cell positioning.

Authors:  Sinwook Park; Dana Ben Bassat; Gilad Yossifon
Journal:  Biomicrofluidics       Date:  2014-04-24       Impact factor: 2.800

4.  Calcein Release from Cells In Vitro via Reversible and Irreversible Electroporation.

Authors:  Violeta Rajeckaitė; Baltramiejus Jakštys; Arnas Rafanavičius; Martynas Maciulevičius; Milda Jakutavičiūtė; Saulius Šatkauskas
Journal:  J Membr Biol       Date:  2017-11-15       Impact factor: 1.843

5.  Microfluidics-Based Chromosome Conformation Capture (3C) Technology for Examining Chromatin Organization with a Low Quantity of Cells.

Authors:  Chen Sun; Chang Lu
Journal:  Anal Chem       Date:  2018-03-08       Impact factor: 6.986

Review 6.  Microfluidic electroporation for cellular analysis and delivery.

Authors:  Tao Geng; Chang Lu
Journal:  Lab Chip       Date:  2013-10-07       Impact factor: 6.799

Review 7.  Microfluidic chip-based technologies: emerging platforms for cancer diagnosis.

Authors:  Li Ying; Qi Wang
Journal:  BMC Biotechnol       Date:  2013-09-27       Impact factor: 2.563

8.  Automated long-term monitoring of parallel microfluidic operations applying a machine vision-assisted positioning method.

Authors:  Hon Ming Yip; John C S Li; Kai Xie; Xin Cui; Agrim Prasad; Qiannan Gao; Chi Chiu Leung; Raymond H W Lam
Journal:  ScientificWorldJournal       Date:  2014-07-15

9.  Focusing of mammalian cells under an ultrahigh pH gradient created by unidirectional electropulsation in a confined microchamber†Electronic supplementary information (ESI) available: Figures S1-S5 and videos S1-S2. See DOI: 10.1039/c4sc00319eClick here for additional data file.Click here for additional data file.Click here for additional data file.

Authors:  Despina Nelie Loufakis; Zhenning Cao; Sai Ma; David Mittelman; Chang Lu
Journal:  Chem Sci       Date:  2014-06-20       Impact factor: 9.825

10.  RNA Extraction from a Mycobacterium under Ultrahigh Electric Field Intensity in a Microfluidic Device.

Authors:  Sai Ma; Bryan D Bryson; Chen Sun; Sarah M Fortune; Chang Lu
Journal:  Anal Chem       Date:  2016-04-27       Impact factor: 6.986

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