دورية أكاديمية
Ultrafast, low-power, PCB manufacturable, continuous-flow microdevice for DNA amplification.
| العنوان: | Ultrafast, low-power, PCB manufacturable, continuous-flow microdevice for DNA amplification. |
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| المؤلفون: | Kaprou GD; Institute of Nanoscience and Nanotechnology, NCSR Demokritos, Patr. Gregoriou E' and 27 Neapoleos Str., PO Box 60037, 15341, Agia Paraskevi, Attica, Greece.; Department of Biology, University of Crete, Voutes, 70013, Heraklion, Greece., Papadopoulos V; Institute of Nanoscience and Nanotechnology, NCSR Demokritos, Patr. Gregoriou E' and 27 Neapoleos Str., PO Box 60037, 15341, Agia Paraskevi, Attica, Greece., Papageorgiou DP; Institute of Nanoscience and Nanotechnology, NCSR Demokritos, Patr. Gregoriou E' and 27 Neapoleos Str., PO Box 60037, 15341, Agia Paraskevi, Attica, Greece.; Department of Materials Science and Engineering, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA., Kefala I; Institute of Nanoscience and Nanotechnology, NCSR Demokritos, Patr. Gregoriou E' and 27 Neapoleos Str., PO Box 60037, 15341, Agia Paraskevi, Attica, Greece., Papadakis G; Institute of Molecular Biology and Biotechnology-FORTH, 100 N. Plastira Str., 70013, Heraklion, Greece., Gizeli E; Department of Biology, University of Crete, Voutes, 70013, Heraklion, Greece.; Institute of Molecular Biology and Biotechnology-FORTH, 100 N. Plastira Str., 70013, Heraklion, Greece., Chatzandroulis S; Institute of Nanoscience and Nanotechnology, NCSR Demokritos, Patr. Gregoriou E' and 27 Neapoleos Str., PO Box 60037, 15341, Agia Paraskevi, Attica, Greece., Kokkoris G; Institute of Nanoscience and Nanotechnology, NCSR Demokritos, Patr. Gregoriou E' and 27 Neapoleos Str., PO Box 60037, 15341, Agia Paraskevi, Attica, Greece. g.kokkoris@inn.demokritos.gr., Tserepi A; Institute of Nanoscience and Nanotechnology, NCSR Demokritos, Patr. Gregoriou E' and 27 Neapoleos Str., PO Box 60037, 15341, Agia Paraskevi, Attica, Greece. a.tserepi@inn.demokritos.gr. |
| المصدر: | Analytical and bioanalytical chemistry [Anal Bioanal Chem] 2019 Aug; Vol. 411 (20), pp. 5297-5307. Date of Electronic Publication: 2019 Jun 03. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: Springer-Verlag Country of Publication: Germany NLM ID: 101134327 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1618-2650 (Electronic) Linking ISSN: 16182642 NLM ISO Abbreviation: Anal Bioanal Chem |
| أسماء مطبوعة: | Original Publication: Heidelberg : Springer-Verlag, 2002- |
| مواضيع طبية MeSH: | Lab-On-A-Chip Devices* , Manufactured Materials*, DNA/*chemistry , Polychlorinated Biphenyls/*chemistry, Polymerase Chain Reaction/methods |
| مستخلص: | The design and fabrication of a continuous-flow μPCR device with very short amplification time and low power consumption are presented. Commercially available, 4-layer printed circuit board (PCB) substrates are employed, with in-house designed yet industrially manufactured embedded Cu micro-resistive heaters lying at very close distance from the microfluidic network, where DNA amplification takes place. The 1.9-m-long microchannel in combination with desirably high flow velocities (for fast amplification) challenged the robustness of the sealing that was overcome with the development of a novel bonding method rendering the microdevice robust even at extreme pressure drops (12 bars). The proposed fabrication methods are PCB compatible, allowing for mass and reliable production of the μPCR device in the established PCB industry. The μPCR chip was successfully validated during the amplification of two different DNA fragments (and with different target DNA copies) corresponding to the exon 20 of the BRCA1 gene, and to the plasmid pBR322, a commonly used cloning vector in E. coli. Successful DNA amplification was demonstrated at total reaction times down to 2 min, with a power consumption of 2.7 W, rendering the presented μPCR one of the fastest and lowest power-consuming devices, suitable for implementation in low-resource settings. Detailed numerical calculations of the DNA residence time distributions, within an acceptable temperature range for denaturation, annealing, and extension, performed for the first time in the literature, provide useful information regarding the actual on-chip PCR protocol and justify the maximum volumetric flow rate for successful DNA amplification. The calculations indicate that the shortest amplification time is achieved when the device is operated at its enzyme kinetic limit (i.e., extension rate). Graphical abstract. |
| معلومات مُعتمدة: | Project ID: 68768 Horizon 2020-EU 2.1.1, "LOVEFOOD2Market- A portable MicroNanoBioSystem and Instrument for ultra-fast analysis of pathogens in food: Innovation from LOVE-FOOD lab prototype to a pre-commercial instrument; Contract No 317742 FP7 "Love Wave Fully Integrated Lab-on-chip Platform for Food Pathogen Detection" - LOVE FOOD project |
| فهرسة مساهمة: | Keywords: Computational fluid dynamics; Continuous-flow; Heat transport; MicroPCR; PCB substrates; Residence time distribution |
| المشرفين على المادة: | 9007-49-2 (DNA) DFC2HB4I0K (Polychlorinated Biphenyls) |
| تواريخ الأحداث: | Date Created: 20190605 Date Completed: 20190730 Latest Revision: 20190730 |
| رمز التحديث: | 20240829 |
| DOI: | 10.1007/s00216-019-01911-1 |
| PMID: | 31161322 |
| قاعدة البيانات: | MEDLINE |
Find this article in full text from Springer Verlag. 
Full Text Finder | تدمد: | 1618-2650 |
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| DOI: | 10.1007/s00216-019-01911-1 |