使用 3D 打印设备浓缩 lambda 连接物。
Concentration of lambda concatemers using a 3D printed device.
机构信息
Department of Chemistry, University of Nebraska-Kearney, Kearney, Nebraska, USA.
出版信息
Electrophoresis. 2023 Apr;44(7-8):744-751. doi: 10.1002/elps.202200200. Epub 2023 Mar 2.
Identifying significant variations in genomes can be cumbersome, as the variations span a multitude of base pairs and can make genome assembly difficult. However, large DNA molecules that span the variation aid in assembly. Due to the DNA molecule's large size, routine molecular biology techniques can break DNA. Therefore, a method is required to concentrate large DNA. A bis-acrylamide roadblock was cured in a proof-of-principle 3D printed device to concentrate DNA at the interface between the roadblock and solution. Lambda concatemer DNA was stained with YOYO-1 and loaded into the 3D printed device. A dynamic range of voltages and acrylamide concentrations were tested to determine how much DNA was concentrated and recovered. The fluorescence of the original solution and the concentrated solution was measured, the recovery was 37% of the original sample, and the volume decreased by a factor of 3 of the original volume.
鉴定基因组中的显著变异可能很繁琐，因为变异跨越了多个碱基对，这使得基因组组装变得困难。然而，跨越变异的大 DNA 分子有助于组装。由于 DNA 分子的体积庞大，常规的分子生物学技术可能会破坏 DNA。因此，需要一种方法来浓缩大的 DNA。在一个原理验证的 3D 打印设备中，双丙烯酰胺障碍物被修复，以在障碍物和溶液之间的界面处浓缩 DNA。用 YOYO-1 对 λ 连接体 DNA 进行染色，并将其装入 3D 打印设备中。测试了一系列电压和丙烯酰胺浓度，以确定浓缩了多少 DNA 以及回收了多少 DNA。测量了原始溶液和浓缩溶液的荧光强度，回收了原始样品的 37%，体积减少了原始体积的 3 倍。
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