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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...

Sanger Sequencing

Sanger Sequencing

DNA sequencing is a fundamental technique that is routinely used in the biological sciences. This method can be applied to a range of questions at different scales - from the sequencing of a cloned DNA fragment or the study of a mutation in a gene up to whole-genome sequencing. However, despite the widespread use of sequencing today, it was not until 1977 that Fredrick Sanger and his collaborators developed the chain-termination method to decode DNA sequences. It relies on the separation of a...

Maxam-Gilbert Sequencing

Maxam-Gilbert Sequencing

In the same year as the discovery of the Sanger sequencing method, another group of scientists, Allan Maxam and Walter Gilbert, demonstrated their chemical-cleavage method for DNA sequencing. The Maxam-Gilbert method relies on using different chemicals that can cleave the DNA sequence at specific sites, the separation of resulting DNA fragments of variable size using electrophoresis, and deciphering the DNA sequence from the resulting gel bands.
Challenges of the Maxam-Gilbert Method
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Related Experiment Video

Updated: Jun 17, 2026

Purifying Plasmid DNA from Bacterial Colonies Using the Qiagen Miniprep Kit

Purifying Plasmid DNA from Bacterial Colonies Using the Qiagen Miniprep Kit

Published on: July 29, 2007

A hot alkaline plasmid DNA miniprep method for automated DNA sequencing protocols

P R Musich¹, W Chu

¹Dept. of Biochemistry, James H. Quillen College of Medicine, East Tennessee State University, Johnson City 37614-0581.

|June 1, 1993

Summary

This study presents a fast, inexpensive hot alkaline lysis method for plasmid DNA isolation. The procedure yields high-quality DNA suitable for automated sequencing and other molecular biology applications.

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Last Updated: Jun 17, 2026

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Published on: August 8, 2019

Area of Science:

Molecular Biology
Biochemistry

Context:

Automated DNA sequencing requires high-quality plasmid DNA substrates.
Current plasmid DNA isolation methods can be time-consuming, expensive, or involve harsh chemicals.

Purpose:

To describe a novel hot alkaline lysis miniprep procedure for plasmid DNA isolation.
To generate high-quality plasmid DNA suitable for automated DNA sequencing and molecular biology applications.

Summary:

A hot alkaline lysis method was developed for plasmid DNA isolation using routine laboratory chemicals.
This enzyme- and organic solvent-free procedure effectively removes RNA contamination.
The method yields >20 micrograms of plasmid DNA from a 5-ml overnight culture.

Impact:

Achieves a >96% success rate in automated DNA sequencing.
Produces plasmid DNA that is compatible with various molecular biology manipulations.
Offers an inexpensive and rapid alternative for plasmid DNA preparation.