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An integrated microfluidic system capable of sample pretreatment and hybridization for microarrays

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Abstract

Sample pretreatment is a critical series of processing steps in most protocols involving microarrays. It usually involves a labor-intensive, time-consuming process, which also requires bulky and costly apparatus to ensure a high-quality RNA extraction. Furthermore, an efficient hybridization process is also critical for practical applications in microarrays. The present study, therefore, reports a new microfluidic system capable of automatically performing the sample pretreatment and hybridization processes for microarrays. Process steps including cell lysis, extraction of messenger ribonucleic acid (mRNA), reverse transcription (RT), purification of complementary deoxyribonucleic acid (cDNA), and hybridization, are all performed automatically in this miniature system. When compared to the conventional methods, which usually require well-trained personnel and time consuming processes, this proposed microfluidic system can perform the entire experimental protocol automatically within 6 h. This developed system may provide a useful platform for subsequent genetic analysis and diagnostic applications.

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Abbreviations

Au:

Gold

BCIP:

5-Bromo-4-chloro3-indolyl-phosphate

CCD:

Charge-coupled device

cDNA:

Complementary deoxyribonucleic acid

CNC:

Computer-numerical-control

DEPC:

Diethyl pyrocarbonate

DI:

Deionized

DIG:

Digoxigenin

dNTP:

Deoxyribonucleotide triphosphate

EtBr:

Ethidium bromide

EMV:

Electromagnetic valve

MEMS:

Micro-electro-mechanical-systems

NBT:

Nitroblue-tetrazolium

mRNA:

Messenger ribonucleic acid

PBS:

Phosphate buffer solution

PCR:

Polymerase chain reaction

PDM:

Spolydimethylsiloxane

PMMA:

Polymethylmethacrylate

RNA:

Ribonucleic acid

RNase:

Ribonuclease

RNAse H:

Ribonuclease H

RT:

Reverse transcription

SNP:

Single nucleotide polymorphism

UV:

Ultraviolet

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Acknowledgments

The authors would like to thank the National Science Council in Taiwan for financial support of this project (NSC 98-2627-B-006-006 and 98-2627-B-006-005). Partial financial support from grant (DOH 98-TD-B-111-004, DOH 99-TD-C-111-003) is also greatly appreciated. The authors also want to extend their appreciation to Professor Shiu-Ru Lin for her assistance on the membrane array.

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Authors and Affiliations

  1. Department of Engineering Science, National Cheng Kung University, Tainan 701, Taiwan

    Chien-Hsuan Tai, Chun-Che Lin & Gwo-Bin Lee

  2. Department of Parasitology, National Cheng Kung University, Tainan 701, Taiwan

    Jyh-Wei Shin & Tsuey-Yu Chang

  3. School of Medical and Health Science, Fooyin University, Kaohsiung 831, Taiwan

    Suz-Kai Hsiung

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  1. Chien-Hsuan Tai
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  2. Jyh-Wei Shin
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  3. Tsuey-Yu Chang
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  4. Suz-Kai Hsiung
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  5. Chun-Che Lin
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  6. Gwo-Bin Lee
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Correspondence to Gwo-Bin Lee.

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Tai, CH., Shin, JW., Chang, TY. et al. An integrated microfluidic system capable of sample pretreatment and hybridization for microarrays. Microfluid Nanofluid 10, 999–1009 (2011). https://doi.org/10.1007/s10404-010-0729-6

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  • DOI: https://doi.org/10.1007/s10404-010-0729-6

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