Name: high-throughput protein expression generator using a microfluidic platform
Uploaded: 2012-08-23T12:00:00
Description: Watch this Scientific Journal Video about High-throughput Protein Expression Generator Using a Microfluidic Platform at JoVE.com

这项工作得到了玛丽·居里国际重返社会补助金。

1。设备制造<ol><li>购买DTPA-D SU-8个控制模具和模具SPR220-7流从斯坦福大学的微流体铸造厂（ <a href="http://www.stanford.edu/group/foundry" target="_blank">www.stanford.edu /组/晶圆代工</a> ）。 </li><li>三甲基氯硅烷（TMCS）蒸汽和10分钟，以促进弹性体释放后的烘烤步骤9公开的硅氧烷模具。 </li><li>基于有机硅弹性体和固化剂（拌匀）在两个不同的比率5:1和20:1的控制和流量模具，分别准备的混合物。的不同?...

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	<li>Maerkl, S. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Integration+column:+Microfluidic+high-throughput+screening.">Integration column: Microfluidic high-throughput screening.</a> Integrative biology quantitative biosciences from nano to macro. 1, 19-29 (2009).</li><li>Hong, J. W., Quake, S. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Integrated+nanoliter+systems.">Integrated nanoliter systems.</a> Nature. 21, 1179-1183 (2003).</li><li>Unger, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Monolithic+Microfabricated+Valves+and+Pumps+by+Multilayer+Soft+Lithography.">A Monolithic Microfabricated Valves and Pumps by Multilayer Soft Lithography.</a> Science. 288, 113-116 (2000).</li><li>Gerber, D., Maerkl, S. J., Quake, S. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+in+vitro+microfluidic+approach+to+generating+protein-interaction+networks.">An in vitro microfluidic approach to generating protein-interaction networks.</a> Nature. 6, 71-74 (2009).</li><li>Einav, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Discovery+of+a+hepatitis+C+target+and+its+pharmacological+inhibitors+by+microfluidic+affinity+analysis.">Discovery of a hepatitis C target and its pharmacological inhibitors by microfluidic affinity analysis.</a> Nature. 26, 1019-1027 (2008).</li><li>Fordyce, P. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=De+novo+identification+and+biophysical+characterization+of+transcription-factor+binding+sites+with+microfluidic+affinity+analysis.">De novo identification and biophysical characterization of transcription-factor binding sites with microfluidic affinity analysis.</a> Nature Biotechnology. 28, 962-967 (2010).</li><li>Zhu, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Global+analysis+of+protein+activities+using+proteome+chips.">Global analysis of protein activities using proteome chips.</a> Science (New York, N.Y.). 293, 2101-2105 (2001).</li><li>Ramachandran, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Self-assembling+protein+microarrays.">Self-assembling protein microarrays.</a> Science (New York, N.Y.). 305, 86-90 (2004).</li><li>Zhong, J. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+microfluidic+processor+for+gene+expression+profiling+of+single+human+embryonic+stem+cells.">A microfluidic processor for gene expression profiling of single human embryonic stem cells.</a> Lab on a chip. 8, 68-74 (2008).</li><li>Kusnezow, W., Hoheisel, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Solid+supports+for+microarray+immunoassays.">Solid supports for microarray immunoassays.</a> Journal of molecular recognition JMR. 16, 165-176 (2003).</li><li>Lundin, M., Monne, M., Widell, A., Von Heijne, G., Persson, M. A. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Topology+of+the+membrane-associated+hepatitis+C+virus+protein+NS4B.">Topology of the membrane-associated hepatitis C virus protein NS4B.</a> Journal of virology. 77, 5428 (2003).</li></ol>

在本文中，我们提出了一个方法代高通量的蛋白质阵列使用微流体平台。上述排列生成是基于微阵列打印的DNA模板， 在体外蛋白质表达的微流体装置内的DNA。 我们的新型微流体平台有几个重要的优点，比目前使用的方法，这使得它有前途和蛋白质组学工具。的一个优点是与膜结合蛋白， 在体外蛋白质合成使用哺乳动物的网织红细胞裂解物在微粒体膜11?...

<table border="1"><tbody><tr><td> 试剂/设备 </td><td> 公司 </td><td> 目录编号 </td></tr><tr><td> PDMS-SYLGARD 184 </td><td>美国道康宁</td><td>的ESSEX-DC </td></tr><tr><td>三甲基氯硅烷（TMCS </td><td> Sigma-Aldrich公司</td><td> C72854 </td></tr><tr><td>环氧涂层的玻璃基板</td><td> CEL Associates公司（美国） </td><td> VEPO-25C </td></tr><tr><td>保利：乙烯glycole（PEG） </td><td> Sigma-Aldrich公司</td><td> 81260 </td></tr><tr><td> D-海藻糖二水合物</td><td> Sigma-Aldrich公司</td><td> T9531 </td></tr><tr><td>生物素-BSA </td><td>刺穿</td><td> PIR-29130 </td></tr><tr><td> Neutravidin </td><td>刺穿</td><td> 31050 </td></tr><tr><td>五-生物素</td><td> Qiagen公司</TD&gt; <td> 34440 </td></tr><tr><td> HEPES </td><td> <a href="http://www.bioind.com/htmls/article.aspx?c0=741&amp;bsp=13384" target="_blank">生物产业</a> </td><td> 03-025-1B </td></tr><tr><td> TNT-T7 </td><td> Promega公司</td><td> L5540 </td></tr><tr><td> C-myc基因Cy3标记抗体</td><td> Sigma-Aldrich公司</td><td></td></tr><tr><td>控制箱</td><td>斯坦福大学的微流体铸造厂</td><td></td></tr><tr><td>模</td><td>斯坦福大学的微流体铸造厂</td><td></td></tr><tr><td>脚</td><td>新英国小管公司</td><td></td></tr><tr><td>聚乙烯微孔管</td><td>聚乙烯</td><td> S-54-HL </td></tr><tr><td>微阵列</td><td>生物机器人</td><td>微网610 </td></tr><tr><td>有机硅针</td><td>并行Syn的论文</td><td> SMT-S75 </td></tr></tbody></table>

high-throughput protein expression generator using a microfluidic platform

迅速增长的领域，如系统生物学，需要制定和实施新技术，使大型系统的高吞吐量和高精确度的测量。 ，微流体承诺履行这些要求的，如片上进行高通量筛选实验，当中包括生物化学，生物物理学，和基于细胞的检测1。微流体设备的初期以来，这一领域的显着发展，领导发展的微流体大规模集成2,3。该技术允许用邮资大小的足迹（ 图1）在单个设备上集成的成千上万的微机械阀。我们已经开发出一种高通量微流体平台， 在体外表达的蛋白质阵列（ 图2）PING（蛋白质相互作用网络发生器）产生。这些阵列可以作为一个模板，为许多实验如蛋白质-蛋白质4 5，蛋白质-RNA或蛋白质-DNA 6相互作用。 该装置由成千上万的反应室，单独使用微阵列编程。对齐的微流体设备方案这些印刷的微阵列，每个腔室与一个单一的点，此外，生成使用标准的微阵列斑点技术的微阵列消除潜在的污染或交叉反应性也非常模块化的，允许的蛋白质7，DNA 8，小分子的排列，甚至胶体悬浮液。微流体生物科学的潜在影响是显著的。已经许多微流控芯片为基础的检测提供了新的见解，对生物系统的结构和功能，和微流控芯片领域将继续影响生物。

Watch this Scientific Journal Video about High-throughput Protein Expression Generator Using a Microfluidic Platform at JoVE.com

High-throughput Protein Expression Generator Using a Microfluidic Platform

我们提出了一个表达的蛋白质阵列微流控方法。该装置由反应室的数以千计的由微机械阀控制。的微流体装置相配合以印刷的微阵列的基因库。然后，这些基因的转录和翻译的片上，所得的蛋白质阵列中准备实验使用。

利用微流控平台的高通量蛋白质表达发生器

Rapidly increasing fields, such as systems biology, require the development and implementation of new technologies, enabling high-throughput and ...

Research

JoVE Journal

Bioengineering

CometChip: A High-throughput 96-Well Platform for Measuring DNA Damage in Microarrayed Human Cells

CometChip：一种高通量的96孔平台的微阵列人类细胞测量DNA损伤

DNA damaging agents can promote aging, disease and cancer and they are ubiquitous in the environment and produced within human cells as normal cellular ...

科研

生物工程

High Throughput Microfluidic Rapid and Low Cost Prototyping Packaging Methods

In this work, 3 different packaging and assembly techniques are presented. They can be classified into two categories: one-time use and reusable packaging ...

Cell Squeezing as a Robust, Microfluidic Intracellular Delivery Platform

电池挤压作为一种强大的，微流控细胞内传送平台

Rapid mechanical deformation of cells has emerged as a promising, vector-free method for intracellular delivery of macromolecules and nanomaterials. This ...

Use of a High-throughput In Vitro Microfluidic System to Develop Oral Multi-species Biofilms

Use of a High-throughput In Vitro Microfluidic System to Develop Oral Multi-species Biofilms

使用高通量的<em&gt;在体外</em&gt;微流控系统开发口服多物种生物膜

There are few high-throughput in vitro systems which facilitate the development of multi-species biofilms that contain numerous species commonly detected ...

A Microfluidic Platform for High-throughput Single-cell Isolation and Culture

高通量单细胞的分离和文化的微流体平台

Studying the heterogeneity of single cells is crucial for many biological questions, but is technically difficult. Thus, there is a need for a simple, yet ...

A High-throughput Cell Microarray Platform for Correlative Analysis of Cell Differentiation and Traction Forces

细胞分化和牵引力的相关分析高通量细胞芯片平台

Microfabricated cellular microarrays, which consist of contact-printed combinations of biomolecules on an elastic hydrogel surface, provide a tightly ...

A Microfluidic Platform for Longitudinal Imaging in Caenorhabditis elegans

A Microfluidic Platform for Longitudinal Imaging in Caenorhabditis elegans

秀丽线虫纵向成像的微流控平台

In the last decade, microfluidic techniques have been applied to study small animals, including the nematode Caenorhabditis elegans, and have proved ...

A Multilayer Microfluidic Platform for the Conduction of Prolonged Cell-Free Gene Expression

用于长期无细胞基因表达传导的多层微流体平台

The limitations of cell-based synthetic biology are becoming increasingly apparent as researchers aim to develop larger and more complex synthetic genetic ...

A Microfluidic Platform for Stimulating Chondrocytes with Dynamic Compression

微流体平台，用于用动态压缩刺激软骨细胞

Mechanical stimuli are known to modulate biological functions of cells and tissues. Recent studies have suggested that compressive stress alters growth ...

High-Throughput Identification of Resistance to Pseudomonas syringae pv. Tomato in Tomato using Seedling Flood Assay

High-Throughput Identification of Resistance to Pseudomonas syringae pv. Tomato in Tomato using Seedling Flood Assay

高通量识别对伪单鱼的电阻syringae pv。番茄中的番茄使用苗子洪水测定

Tomato is an agronomically important crop that can be infected by Pseudomonas syringae, a Gram-negative bacterium, resulting in bacterial speck disease. ...