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DNA as a Genetic Template

DNA as a Genetic Template

Two structural features of the DNA molecule provide a basis for the mechanisms of heredity: the four nucleotide bases and its double-stranded nature. The Watson-Crick model of double-helical DNA structure, proposed in 1952, drew heavily upon the X-ray crystallography work of researchers Rosalind Franklin and Maurice Wilkins. Watson, Crick, and Wilkins jointly received the Nobel Prize in Physiology or Medicine for their work in 1962. Franklin was, controversially, excluded from the prize for...

DNA as a Genetic Template

DNA as a Genetic Template

Two structural features of the DNA molecule provide a basis for the mechanisms of heredity: the four nucleotide bases and its double-stranded nature. The Watson-Crick model of double-helical DNA structure, proposed in 1952, drew heavily upon the X-ray crystallography work of researchers Rosalind Franklin and Maurice Wilkins. Watson, Crick, and Wilkins jointly received the Nobel Prize in Physiology or Medicine for their work in 1962. Franklin was, controversially, excluded from the prize for...

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Related Experiment Video

Updated: Jul 14, 2026

Automated Robotic Liquid Handling Assembly of Modular DNA Devices

Automated Robotic Liquid Handling Assembly of Modular DNA Devices

Published on: December 1, 2017

Modular Golden Gate Assembly of Linear DNA Templates for Cell-Free Prototyping.

François-Xavier Lehr^1,2, Aukse Gaizauskaite^1,2,3, Katarzyna Elżbieta Lipińska^1,2

¹Max Planck Institute for Terrestrial Microbiology, Marburg, Germany.

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

|October 3, 2024

Summary

This study presents a rapid, cloning-free method for creating DNA templates for cell-free transcription and translation (TXTL) systems. This accelerates synthetic biology prototyping by using Golden Gate assembly for faster genetic circuit testing.

Keywords:

DNA assembly E. coli cell-free systems Golden Gate cloning In vitro transcription translation Linear DNA Rapid prototyping

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Rapid Assembly of Multi-Gene Constructs using Modular Golden Gate Cloning

Rapid Assembly of Multi-Gene Constructs using Modular Golden Gate Cloning

Published on: February 5, 2021

Area of Science:

Synthetic Biology
Molecular Biology
Biotechnology

Background:

Cell-free transcription and translation (TXTL) systems are valuable for genetic circuit testing.
Accelerating the design-build-test-learn cycle is crucial in synthetic biology.
Current methods often involve time-consuming cellular cloning steps.

Purpose of the Study:

To develop a rapid, cloning-free workflow for producing linear DNA templates for TXTL.
To streamline the assembly and testing of genetic regulatory elements.
To enhance the efficiency of synthetic biology prototyping.

Main Methods:

Utilized Golden Gate assembly for in vitro construction of transcription units.
Assembled DNA templates from basic genetic parts (promoter, RBS, coding sequence, terminator).
Employed polymerase chain reaction (PCR) amplification for template production.

Main Results:

Successfully demonstrated the assembly of functional DNA templates.
Showcased cell-free testing of various promoter and ribosomal binding site (RBS) combinations.
Characterized a repressor-promoter genetic pair using the developed workflow.

Conclusions:

The described workflow significantly accelerates the prototyping of synthetic biology circuits.
Eliminating cellular transformation and cloning steps saves considerable time.
This method enables rapid generation of data for numerous genetic constructs within a day.