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DNA Isolation

DNA Isolation

DNA isolation protocols can be fast and straightforward or complex and time-consuming depending on the type and quality of DNA required for further processing. For example, plasmid DNA extraction is a bit more complicated than genomic DNA extraction because of the need for an appropriate lysis method to separate plasmid DNA from gDNA during isolation. However, for specific applications, such as long-range DNA sequencing that require a good yield of high- quality DNA samples, we need to follow...

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Updated: Jun 14, 2025

Production of Double-stranded DNA Ministrings

Production of Double-stranded DNA Ministrings

Published on: February 29, 2016

为高效的大型DNA在微生物之间提供无需 in vitro 净化的多功能方法.

Shinya Kaneko¹, Hiromi Fukushima¹, Misako Nakahama¹

¹School of Life Science and Technology, Institute of Science Tokyo, Yokohama, Kanagawa 226-8501, Japan.

Journal of molecular biology

|June 12, 2025

概括

来自溶解细菌的细胞外等离子体可以在不经净化的情况下转化大肠杆菌和大脑菌. 这种细胞溶解技术提供可变细胞外DNA (CELyTED) 方法,有效地提供大型DNA质粒.

关键词:

细胞溶解是细胞的溶解.细胞外DNA是细胞外DNA.基因组合成的方法横向的基因转移是可以实现的.变化转化转化转化

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相关实验视频

Last Updated: Jun 14, 2025

Production of Double-stranded DNA Ministrings

Production of Double-stranded DNA Ministrings

Published on: February 29, 2016

Microscale Vortex-assisted Electroporator for Sequential Molecular Delivery

Microscale Vortex-assisted Electroporator for Sequential Molecular Delivery

Published on: August 7, 2014

High-Throughput DNA Plasmid Multiplexing and Transfection Using Acoustic Nanodispensing Technology

High-Throughput DNA Plasmid Multiplexing and Transfection Using Acoustic Nanodispensing Technology

Published on: August 8, 2019

科学领域:

微生物学微生物学
分子生物学分子生物学
合成生物学合成生物学

背景情况:

传统的微生物转化依赖于纯化的DNA等离子体.
净化过程可以切割大型DNA分子,限制它们的实用性.
需要简化,高效的DNA输送方法.

研究的目的:

引入一种使用细胞外等离子体的替代DNA输送方法.
为了证明细胞溶解技术的有效性,提供可转换的细胞外DNA (CELyTED) 协议.
为了使CELYTED适应转化原生和真核微生物.

主要方法:

使用了从溶解的 Bacillus subtilis 中释放的细胞外质粒.
应用CELYTED用于直接转化具有化学能力的大肠杆菌.
适应了Saccharomyces cerevisiae转化的协议.
优化了供体宿主细胞溶解的条件.

主要成果:

通过溶解的供体宿主细胞成功转化了大肠杆菌和大肠杆菌.
通过CELyTED,可以提供超过50kB的DNA质粒.
该方法绕过了对生物化学DNA净化的需求.
对于 prokaryotic 和 eukaryotic 微生物均表现出多功能性.

结论:

CELYTED提供了一种简化的基因转换方法.
这项技术有助于引入大型DNA质粒,保持它们的完整性.
CELyTED为合成基因组应用提供了一个基础平台.
这种方法简化了DNA传递,提高了其在各种微生物系统中的适用性.