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PCR

PCR

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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Development of a Quantitative Recombinase Polymerase Amplification Assay with an Internal Positive Control

Development of a Quantitative Recombinase Polymerase Amplification Assay with an Internal Positive Control

Published on: March 30, 2015

Colony PCR.

Flávio Azevedo¹, Humberto Pereira¹, Björn Johansson²

¹Centre of Molecular and Environmental Biology (CBMA), Department of Biology, University of Minho, Campus de Gualtar, 4710-057, Braga, Portugal.

Methods in Molecular Biology (Clifton, N.J.)

|May 26, 2017

Summary

This summary is machine-generated.

This study presents efficient colony PCR protocols for Escherichia coli and Saccharomyces cerevisiae. These methods simplify DNA verification in synthetic biology, overcoming challenges with DNA release and PCR inhibitors.

Keywords:

Colony Direct lysis Escherichia coli PCR Saccharomyces cerevisiae Yeast

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Last Updated: Nov 17, 2025

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Area of Science:

Synthetic biology
Molecular biology
Microbiology

Background:

Escherichia coli and Saccharomyces cerevisiae are key organisms in synthetic biology.
Yeast is crucial for eukaryotic gene expression studies and DNA assembly.
Verifying DNA sequences via colony PCR is essential but challenging due to DNA release and inhibitor issues.

Purpose of the Study:

To develop and present efficient colony PCR protocols for E. coli and S. cerevisiae.
To address the limitations of current colony PCR methods, including DNA release and inhibitor presence.
To provide an overview of past and future developments in yeast colony PCR.

Main Methods:

Developed one protocol for E. coli colony PCR.
Developed two distinct protocols for S. cerevisiae colony PCR, varying in efficiency and complexity.
Reviewed existing and potential future advancements in S. cerevisiae colony PCR.

Main Results:

Presented novel colony PCR protocols for both E. coli and S. cerevisiae.
Protocols aim to improve DNA release and mitigate PCR inhibitors.
Offered insights into protocol efficiency and complexity trade-offs.

Conclusions:

The developed protocols offer improved methods for DNA sequence verification in synthetic biology.
These advancements facilitate faster screening in E. coli and S. cerevisiae based research.
The study contributes to the ongoing development of efficient molecular biology techniques.