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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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Related Experiment Video

Updated: Jun 14, 2025

Production of Double-stranded DNA Ministrings

Production of Double-stranded DNA Ministrings

Published on: February 29, 2016

Versatile Methodology for Efficient Large-sized DNA Delivery Between Microorganisms Without In vitro Purification.

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

Summary

Extracellular plasmids from lysed bacteria can transform E. coli and S. cerevisiae without purification. This Cell Lysis Technology to provide Transformable Extra-cellular DNA (CELyTED) method efficiently delivers large DNA plasmids.

Keywords:

cell lysis extracellular DNA genome synthesis horizontal gene transfer transformation

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Last Updated: Jun 14, 2025

Production of Double-stranded DNA Ministrings

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Published on: February 29, 2016

Microscale Vortex-assisted Electroporator for Sequential Molecular Delivery

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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

Area of Science:

Microbiology
Molecular Biology
Synthetic Biology

Background:

Traditional microbial transformation relies on purified DNA plasmids.
Purification processes can shear large DNA molecules, limiting their utility.
A need exists for simplified, efficient DNA delivery methods.

Purpose of the Study:

To introduce an alternative DNA delivery method using extracellular plasmids.
To demonstrate the efficacy of the Cell Lysis Technology to provide Transformable Extra-cellular DNA (CELyTED) protocol.
To adapt CELyTED for transforming both prokaryotic and eukaryotic microorganisms.

Main Methods:

Utilized extracellular plasmids released from lysed Bacillus subtilis.
Applied CELyTED for direct transformation of chemically competent Escherichia coli.
Adapted the protocol for Saccharomyces cerevisiae transformation.
Optimized conditions for donor host cell lysis.

Main Results:

Successfully transformed E. coli and S. cerevisiae using lysed donor host cells.
CELyTED enabled the delivery of DNA plasmids larger than 50 kb.
The method bypassed the need for biochemical DNA purification.
Demonstrated versatility for both prokaryotic and eukaryotic microorganisms.

Conclusions:

CELyTED offers a streamlined approach for genetic transformation.
The technology facilitates the introduction of large DNA plasmids, preserving their integrity.
CELyTED presents a foundational platform for synthetic genome applications.
This method simplifies DNA delivery, enhancing its applicability in diverse microbial systems.