Name: changing the direction and orientation of electric field during electric pulses application improves plasmid gene transfer in vitro
Uploaded: 2011-09-12T14:00:00
Description: Watch this Scientific Journal Video about Changing the Direction and Orientation of Electric Field During Electric Pulses Application Improves Plasmid Gene Transfer in vitro at JoVE.com

这项工作得到了斯洛文尼亚的研究机构（项目，基建中心IP J2 - 9770 - 0510和程序P2 - 0249）。这个视频代表的“电穿孔技术为基础的技术和处理”的科学研讨会，并在斯洛文尼亚的卢布尔雅那大学电气工程学院举办的研究生课程，补充材料。作者还感谢Duša Hodžič请提供质粒DNA。

1。细胞培养，质粒和缓冲区，实验的准备<ol><li>在这个实验中，使用中国仓鼠卵巢细胞（CHO - K1）。细胞是生长在一种营养混合物的HAM - F12辅以2毫米L -谷氨酰胺，10％胎牛血清，400μL/ L庆大霉素（所有自Sigma - Aldrich化学公司，Deisenhofen，德国），和1（PAA）的毫升/升crystacilin（Pliva公司，萨格勒布，克罗地亚）。细胞都保持在37 ° C，饱和湿度5％的CO 2，在孵化器的气氛为24小时。 </li><li>放?...

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基因electrotransfer是一个多功能的生物科技技术，使进入细胞的DNA转移，通过应用短，高电压电脉冲，并由于其安全性，有效性和易于应用的病毒载体，代表了更安全的替代。虽然在今天的基因electrotransfer被广泛用于转染所有类型的细胞和第一阶段的临床试验，使用这种方法，已报道4，底层的机制仍然没有完全理解。据了解，应用足够的实力来的细胞的电脉冲，导致跨膜电位的增加，从而?...

<table border="1"><tbody><tr><td> 试剂名称 </td><td> 公司 </td><td> 目录编号 </td><td> 评论 </td></tr><tr><td> HAM - F12 </td><td>临时机场管理局</td><td> E15 - 016 </td><td>培养基</td></tr><tr><td> L -谷氨酰胺</td><td> Sigma - Aldrich公司</td><td> G7513 </td><td></td></tr><tr><td>胎牛血清</td><td>临时机场管理局</td><td> A15 - 151 </td><td></td></tr><tr><td>庆大霉素</td><td> Sigma - Aldrich公司</td><td> G1397 </td><td>抗生素</td></tr><tr><td> crystacilin </td><td>普利瓦</td><td> 625110 </td><td>抗生素</td></tr><tr><td>以pEGFP - N1 </td><td> Clontech公司实验室</td><td> 6085-1 </td><td>质粒DNA </td></tr><tr><td> HiSpeed​​质粒马克西套件</td><td> Qiagen公司</td><td> 12662 </td><td></td></tr><tr><td> NA 2 HPO 4 </td><td>默克公司</td><td> F640786 933 </td><td></td></tr><tr><td>的NaH 2 PO 4 </td><td> TKI的Hrastnik </td><td> 0795 </td><td></td></tr><tr><td> MgCl 2的 </td><td> Sigma - Aldrich公司</td><td> M - 8266 </td><td></td></tr><tr><td>蔗糖</td><td> Sigma - Aldrich公司</td><td> 16104 </td><td></td></tr><tr><td>胰蛋白酶/ EDTA溶液</td><td> Sigma - Aldrich公司</td><td> T4174 </td><td></td></tr><tr><td>枪头</td><td>定制</td><td></td><td></td></tr><tr><td>电脉冲发生器</td><td>定制</td><td></td><td></td></tr><tr><td> 6孔板</td><td> TPP </td><td> 92406 </td><td></td></tr><tr><td> 15 mL离心管</td><td> TPP </td><td> 91015 </td><td></td></tr><tr><td> 75厘米的培养瓶中</td><td> TPP </td><td> 90076 </td><td></td></tr></tbody></table>

changing the direction and orientation of electric field during electric pulses application improves plasmid gene transfer in vitro

基因electrotransfer是一种物理方法，用于提供短期和强烈的电脉冲，从而导致细胞膜不稳定的应用进入细胞的基因，使其渗透到小分子，并允许转让大分子如DNA。它代表了一个病毒载体，由于其安全性，有效性和易于应用。对于基因electrotransfer使用不同的电脉冲协议，以达到最大的基因转染，其中之一是在脉冲传递改变电场的方向和方位。改变电场方向和方向，增加主管DNA进入细胞的膜面积。在这段视频中，我们展示了基因electrotransfer疗效差异时所有的脉冲是在同一个方向，并交付，脉冲时，通过改变或者电场方向和方向交付。对于这个集成了电极和高电压的原型生成器，它允许在电脉冲的应用不同方向的电场变化的目的端，分别使用。基因electrotransfer疗效是确定的脉冲数量除以所有细胞表达绿色荧光蛋白与细胞的数量的应用后的24小时。结果表明，基因转染增加时，在电脉冲传递的电场方向改变。

Watch this Scientific Journal Video about Changing the Direction and Orientation of Electric Field During Electric Pulses Application Improves Plasmid Gene Transfer in vitro at JoVE.com

Changing the Direction and Orientation of Electric Field During Electric Pulses Application Improves Plasmid Gene Transfer in vitro

基因转染，电是提高约两​​次脉冲的应用过程中改变电场方向时，细胞活力不受影响。基因转染的增加是由主管DNA进入细胞的膜面积增加。

的方向转变，并在电脉冲电场方向的应用，提高质粒的基因转移在体外</em

Gene electrotransfer is a physical method used to deliver genes into the cells by application of short and intense electric pulses, which cause ...

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Changing the Direction and Orientation of Electric Field During Electric Pulses Application Improves Plasmid Gene Transfer in vitro

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