开发一种插入物,在细胞裂解过程中保护大的DNA分子。

IF 2.2 4区 工程技术 Q3 BIOCHEMICAL RESEARCH METHODS
Charles Polen, Esmeralda Mendez Ortiz, Cordelia Harbison, Nate Garringer, Martin Lopez, Kristy L Kounovsky-Shafer
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引用次数: 0

摘要

如果没有长分子来帮助基因组组装,确定大的结构变异是困难和耗时的。在常规的分子生物学技术中,一个问题是大的DNA分子的脆弱性。一种新的倒置琼脂糖插入物被创造出来,以保护和更容易地访问大的DNA。倒置的琼脂糖插入物在中间是细胞溶液,琼脂糖在外面,而不是嵌入在琼脂糖中的细胞(插塞插入物)。测试了多种琼脂糖浓度,以确定创建倒置插入物的琼脂糖的最佳百分比。倒置插入物的原理验证用酿酒酵母细胞进行了测试,结果表明细胞可以在倒置插入物内裂解,使DNA在中间保持全长。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Developing an insert to protect large DNA molecules during cell lysis.

Determining large structural variations is difficult and time-consuming without long molecules to aid in the genome assembly. One issue is the fragility of large DNA molecules during routine molecular biology techniques. A novel inverted agarose insert was created to protect and make accessing large DNA easier. The inverted agarose insert has the cell solution in the middle and the agarose on the outside rather than the cells embedded in the agarose (plug inserts). Multiple agarose concentrations were tested to determine the best percentage of agarose to create the inverted insert. A proof-of-principle inverted insert was tested with S. cerevisiae cells to show that cells could be lysed inside the inverted insert, allowing the DNA to remain at full length in the middle.

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来源期刊
BioTechniques
BioTechniques 工程技术-生化研究方法
CiteScore
4.40
自引率
0.00%
发文量
68
审稿时长
3.3 months
期刊介绍: BioTechniques is a peer-reviewed, open-access journal dedicated to publishing original laboratory methods, related technical and software tools, and methods-oriented review articles that are of broad interest to professional life scientists, as well as to scientists from other disciplines (e.g., chemistry, physics, computer science, plant and agricultural science and climate science) interested in life science applications for their technologies. Since 1983, BioTechniques has been a leading peer-reviewed journal for methods-related research. The journal considers: Reports describing innovative new methods, platforms and software, substantive modifications to existing methods, or innovative applications of existing methods, techniques & tools to new models or scientific questions Descriptions of technical tools that facilitate the design or performance of experiments or data analysis, such as software and simple laboratory devices Surveys of technical approaches related to broad fields of research Reviews discussing advancements in techniques and methods related to broad fields of research Letters to the Editor and Expert Opinions highlighting interesting observations or cautionary tales concerning experimental design, methodology or analysis.
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