Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

Genome Annotation and Assembly

Genome Annotation and Assembly

The genome refers to all of the genetic material in an organism. It can range from a few million base pairs in microbial cells to several billion base pairs in many eukaryotic organisms. Genome assembly refers to the process of taking the DNA sequencing data and putting it all back together in a correct order to create a close representation of the original genome. This is followed by the identification of functional elements on the newly assembled genome, a process called genome annotation.

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...

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by

Same author

Cytokinin-mediated trichome initiation in Nicotiana benthamiana following Agrobacterium tumefaciens infiltration.

Plant communications·2026

Same author

An A. thaliana mutant lacking all nine ATG8 isoforms provides genetic evidence for functional specialization of ATG8 in plants.

Journal of cell science·2025

Same author

Modularization of the type II secretion gene cluster from Xanthomonas euvesicatoria facilitates the identification of a structurally conserved XpsCLM assembly platform complex.

PLoS pathogens·2025

Same author

The suberin transporter StABCG1 is required for barrier formation in potato leaves.

Scientific reports·2025

Same author

Golden Gate Cloning of MoClo Standard Parts.

Methods in molecular biology (Clifton, N.J.)·2024

Same author

Golden Gate Cloning of Multigene Constructs Using the Modular Cloning System MoClo.

Methods in molecular biology (Clifton, N.J.)·2024

Search research articles

Related Experiment Video

Updated: May 8, 2026

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

Combinatorial DNA assembly using Golden Gate cloning.

Carola Engler¹, Sylvestre Marillonnet

¹NOMAD BIOSCIENCE GMBH, Weinbergweg 22, Halle (Saale), Germany.

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

|September 3, 2013

Summary

This summary is machine-generated.

Golden Gate cloning enables efficient DNA assembly for synthetic biology. This one-tube method facilitates parallel assembly of multiple DNA fragments, crucial for constructing complex genetic elements and performing DNA shuffling.

More Related Videos

Genetic Modification of Cyanobacteria by Conjugation Using the CyanoGate Modular Cloning Toolkit

Genetic Modification of Cyanobacteria by Conjugation Using the CyanoGate Modular Cloning Toolkit

Published on: October 31, 2019

Standardized Modular Assembly of Polycistronic Operons with Modular Cloning (MoClo) using the In-Cloning toolkit

Standardized Modular Assembly of Polycistronic Operons with Modular Cloning (MoClo) using the In-Cloning toolkit

Published on: September 2, 2025

Related Experiment Videos

Last Updated: May 8, 2026

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

Genetic Modification of Cyanobacteria by Conjugation Using the CyanoGate Modular Cloning Toolkit

Genetic Modification of Cyanobacteria by Conjugation Using the CyanoGate Modular Cloning Toolkit

Published on: October 31, 2019

Standardized Modular Assembly of Polycistronic Operons with Modular Cloning (MoClo) using the In-Cloning toolkit

Standardized Modular Assembly of Polycistronic Operons with Modular Cloning (MoClo) using the In-Cloning toolkit

Published on: September 2, 2025

Area of Science:

Synthetic Biology
Molecular Biology
Biotechnology

Background:

Efficient DNA assembly is fundamental for synthetic biology applications.
Golden Gate cloning offers a versatile one-tube method for parallel assembly of DNA fragments.
Previous work established Golden Gate cloning for various construct levels.

Purpose of the Study:

To provide a detailed protocol for DNA assembly using Golden Gate cloning.
To exemplify the assembly of gene fragments into complete coding sequences for DNA shuffling.
To outline a standardized method for constructing complex DNA molecules.

Main Methods:

Selection of fusion sites within parental DNA sequences.
Amplification of DNA fragments using PCR with added flanking restriction sites.
Cloning of amplified fragments into intermediate constructs.
One-pot restriction-ligation assembly of intermediate constructs into a recipient vector.

Main Results:

A step-by-step protocol for Golden Gate DNA assembly is presented.
The method effectively assembles gene fragments into complete coding sequences.
This protocol supports applications such as DNA shuffling for protein engineering.

Conclusions:

Golden Gate cloning provides an efficient and versatile platform for DNA assembly in synthetic biology.
The presented protocol simplifies the construction of complex DNA constructs, including gene fusions.
This method is valuable for researchers aiming to engineer genetic systems and perform directed evolution.