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Reproductive Cloning

Reproductive Cloning

Reproductive cloning is the process of producing a genetically identical copy—a clone—of an entire organism. While clones can be produced by splitting an early embryo—similar to what happens naturally with identical twins—cloning of adult animals is usually done by a process called somatic cell nuclear transfer (SCNT).
Somatic Cell Nuclear Transfer
In SCNT, an egg cell is taken from an animal and its nucleus is removed, creating an enucleated egg. Then a somatic cell—any cell that is not a sex...

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CRISPR-based Shuttle Cloning: A High-throughput Cloning Method

CRISPR-based Shuttle Cloning: A High-throughput Cloning Method

Published on: June 13, 2025

A one pot, one step, precision cloning method with high throughput capability.

Carola Engler¹, Romy Kandzia, Sylvestre Marillonnet

¹Icon Genetics GmbH, Biozentrum Halle, Halle, Germany.

|November 6, 2008

Summary

This summary is machine-generated.

Golden Gate cloning offers a precise method for DNA fragment transfer, avoiding unwanted sequences in the final construct. This efficient technique enhances genetic manipulation by eliminating extra amino acids from expressed proteins.

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Subcloning Plus Insertion (SPI) - A Novel Recombineering Method for the Rapid Construction of Gene Targeting Vectors

Published on: January 8, 2015

Area of Science:

Molecular Biology
Genetic Engineering

Background:

Site-specific recombination cloning is efficient but leaves unwanted sequences.
These sequences add extra amino acids to expressed proteins, impacting precision.

Purpose of the Study:

To develop a subcloning strategy that transfers DNA fragments without leaving unwanted sequences.
To provide a more precise method for genetic manipulation.

Main Methods:

Utilized type IIs restriction enzymes that cut outside their recognition sites.
Developed a Golden Gate cloning strategy for one-step restriction-ligation.

Main Results:

Achieved nearly 100% correct recombinant plasmids.
The process is rapid, with restriction-ligation completed in 5 minutes.
Resulting plasmids lack unwanted recombination site sequences.

Conclusions:

Golden Gate cloning is as efficient as recombination-based methods.
This technique offers enhanced precision by yielding constructs without extraneous sequences.
Provides a valuable tool for fundamental genetic manipulation processes.