Name: real-time tracking of dna fragment separation by smartphone
Uploaded: 2017-06-01T15:00:00
Description: Watch this Scientific Journal Video about Real-time Tracking of DNA Fragment Separation by Smartphone at JoVE.com

我们非常感谢中国国家自然科学基金（21205078）和中国高等教育博士研究基金（No.20123120110002）的支持。这项工作得到了中国国家重点研究发展计划（2016YFB1102303），中国国家基础研究计划（973计划，2015CB352001）和国家自然科学基金（61378060）的部分支持。

SGE芯片的基本设计<ol><li>使用任何透明塑料，如聚甲基丙烯酸甲酯（PMMA）或聚碳酸酯。 注：SGE芯片如图1B所示 。 SGE芯片由用于TBE缓冲器的圆柱形孔，用于DNA分离的通道和沿着电极孔嵌入的两条通道组成。 </li><li>使用激光雕刻机在PMMA块中制造SGE通道阵列。 注意：芯片的几何参数取决于要分离的DNA的大小。例如，如果DNA片段小于1,000bp，则SGE芯片的最佳信道参数为2...

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琼脂糖凝胶电泳广泛用于分离DNA，RNA和蛋白质。这项工作提出了一种替代传统凝胶电泳方法的新方法。结果表明，在这样一个小的组装装置中，50,100和1,000bp的DNA梯子可以很好地分离。该方法的优点在于不仅可以将化学消耗少的核酸分开，还可以记录分离过程。尽管DNA片段在图2中看起来很宽，但可以通过优化SGE芯片的参数和琼脂糖凝胶的浓度来改善结果。此外，如果在芯片?...

<html>板状凝胶电泳（SGE）是DNA片段分离最有效的方法1,2,3,4,5 ，因此被认为是生物化学和生物学分析中的通用工具6,7,8。然而，许多实验表明，SGE受到以下四个问题的限制：（1）分离需要很多小时甚至几天; （2）化学品消耗量非常高; （3）需要复杂的装置（ 例如， 2D电泳池，电泳电源和凝胶成像系统）; （4）实验完成后，凝胶成像系统只能观察分离的DNA片段。此外，SGE 9中通常使用的溴化乙锭（EtBr）ass =“xref”&gt; 10，是致突变性和致癌性11,12 。因此，在装载含有EtBr的凝胶时，应始终佩戴手套。 与SGE相比，毛细管电泳（CE）具有许多优点，如自动操作，分离时间短，消耗量少等优点，13，14，15，16，17。然而，CE仪器相当昂贵。因此，为了克服这些限制，已经开发了一种用于分离DNA的系统（ 图1 ）。这样的系统不仅可以大大降低化学消耗，还可以节省SGE实验时间（&lt;8分钟），而且可以通过智能手机对琼脂糖凝胶中的DNA分离过程进行实时跟踪。按照本协议中描述的程序，学生们可以可以设计和制造SGE芯片，在芯片中制备琼脂糖凝胶，用智能手机设置简单的SGE系统，并将DNA迁移过程记录在琼脂糖凝胶中。

<table border="0" cellpadding="0" cellspacing="0" width="683">
	<tbody>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">10&#215;TBE</td>
			<td style="width:287px;">Beijing Solarbio Science &#38; Technology Co., Ltd.</td>
			<td style="width:181px;">T1051</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">50bp DNA ladder</td>
			<td style="width:287px;">Takara Bio Inc.</td>
			<td style="width:181px;">3421A</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">100bp DNA ladder</td>
			<td style="width:287px;">Takara Bio Inc.</td>
			<td style="width:181px;">3422A</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">1kbp DNA ladder</td>
			<td style="width:287px;">Takara Bio Inc.</td>
			<td style="width:181px;">3426A</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">SYBR GREEN</td>
			<td style="width:287px;">Takara Bio Inc.</td>
			<td style="width:181px;">5760A</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Agarose</td>
			<td style="width:287px;">Sigma-Aldrich Corporate</td>
			<td style="width:181px;">V900510</td>
		</tr>
	</tbody>
</table>

real-time tracking of dna fragment separation by smartphone

板状凝胶电泳（SGE）是分离DNA片段的最常用方法;因此，它广泛应用于生物学等领域。然而，传统的SGE协议相当乏味，实验需要很长时间。此外，SGE实验中的化学消耗量非常高。这项工作提出了一种基于SGE芯片分离DNA片段的简单方法。该芯片由雕刻机制成。两个塑料片用于光信号的激发和发射波长。通过智能手机收集DNA条带的荧光信号。为了验证该方法，分离出50,100和1,000bp DNA梯。结果表明，使用该方法可以在12分钟内解析出小于5,000bp的DNA梯度，分辨率高，表明它是传统SGE方法的理想替代品。

<html> 图4 ， 图5和图6代表了在50,100和1,000bp DNA梯级的凝胶电泳之后的典型结果。实验后，DNA片段分离良好。此外，在SGE芯片的4个通道中分离相同的样品，显示出相同尺寸的DNA片段在每个实验中移动相同的距离。 DNA梯度的分离性能可以通过分析迁移距离与DNA大小对数的关系进行评估;因此，SGE可用于计算未知DNA...

Watch this Scientific Journal Video about Real-time Tracking of DNA Fragment Separation by Smartphone at JoVE.com

Real-time Tracking of DNA Fragment Separation by Smartphone

通过智能手机实时跟踪DNA片段分离

传统的平板凝胶电泳（SGE）实验需要复杂的装置和高的化学消耗。这项工作提出了一个协议，描述了一种在短时间内分离DNA片段的低成本方法。

Slab gel electrophoresis (SGE) is the most common method for the separation of DNA fragments; thus, it is broadly applied to the field of biology and ...

Research

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We have developed a method to measure binding of adenine nucleotides to intact, functional transmembrane receptors in a cellular or membrane environment. ...

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Fast photochemical oxidation of proteins (FPOP) is a mass spectrometry-based structural biology technique that probes the solvent-accessible surface area ...

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