New to Cloning?

There are several methods and techniques available for cloning. Traditionally, cloning has utilized restriction enzymes to excise the DNA of interest, and to linearize a plasmid vector while creating compatible ends. After purification of the insert and vector, both are joined with the activity of a DNA ligase, and the newly-created recombinant vector is used to transform an E. coli host for propagation. PCR has also been used to generate both the vector and insert, which can be joined using a variety of techniques, ranging from standard DNA ligation or enzymatic joining using a recombinase or topoisomerase, to homologous recombination. More recently, DNA assembly techniques have been developed that utilize a mixture of enzymes to assemble DNA fragments in a single tube.

Learn more about the cloning workflow

 

Whether you are new to cloning, or having difficulties with an existing experiment, NEB offers a wide selection products, tools and resources that can help you be more efficient and successful with your experiments. To get started, choose the step in the cloning workflow below that you are interested in to find recommended products, videos, technical tips and more.

 

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DNA Assembly

Ordered assembly of multiple DNA fragments to create larger DNA structures has facilitated the field of synthetic biology. This application lends itself well to cloning of large or multiple fragments into a single vector. This technique is rapid and efficient, and can often be performed in a single tube.

 
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Nucleic acid purification

Purification of nucleic acids is an important part of the cloning workflow. Once plasmids containing a desired gene of interest are generated, they can be propagated using competent cells to increase the quantity of desired DNA. Plasmids then need to be recovered from their bacterial hosts using plasmid purification methods. It is important to separate plasmid DNA from the RNA and genomic DNA contained in host cells. Further, DNA must be concentrated and free from contaminating salts to achieve optimal enzyme activity. NEB’s Monarch® Nucleic Acid Purification kits enable quick and easy purification of high quality DNA for direct use in a variety of downstream applications.

 
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Restriction enzyme digestion

Ordered assembly of multiple DNA fragments to create larger DNA structures has facilitated the field of synthetic biology. This application lends itself well to cloning of large or multiple fragments into a single vector. This technique is rapid and efficient, and can often be performed in a single tube.

 
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PCR/amplification

Ordered assembly of multiple DNA fragments to create larger DNA structures has facilitated the field of synthetic biology. This application lends itself well to cloning of large or multiple fragments into a single vector. This technique is rapid and efficient, and can often be performed in a single tube.

 
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End modification

Ordered assembly of multiple DNA fragments to create larger DNA structures has facilitated the field of synthetic biology. This application lends itself well to cloning of large or multiple fragments into a single vector. This technique is rapid and efficient, and can often be performed in a single tube.

 
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DNA ligation

Ordered assembly of multiple DNA fragments to create larger DNA structures has facilitated the field of synthetic biology. This application lends itself well to cloning of large or multiple fragments into a single vector. This technique is rapid and efficient, and can often be performed in a single tube.

 
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Transformation

Ordered assembly of multiple DNA fragments to create larger DNA structures has facilitated the field of synthetic biology. This application lends itself well to cloning of large or multiple fragments into a single vector. This technique is rapid and efficient, and can often be performed in a single tube.

 
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DNA analysis

Ordered assembly of multiple DNA fragments to create larger DNA structures has facilitated the field of synthetic biology. This application lends itself well to cloning of large or multiple fragments into a single vector. This technique is rapid and efficient, and can often be performed in a single tube.

One or more of these products are covered by patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc. For more information, please email us at gbd@neb.com. The use of these products may require you to obtain additional third party intellectual property rights for certain applications.

Feature Cloning Resources

ONLINE RESOURCES

NEBNext_portal_web_icon  Getting Started with Molecular Cloning
Explore simple tips to improve efficiency in your cloning experiment.
NEBNext_portal_web_icon  Troubleshooting Guide for Cloning
Find common problems and solutions associated with cloning experiments.
NEBNext_portal_web_icon  Traditional Cloning Quick Guide
Find quick tips for optimization of each step in the cloning workflow.

ONLINE TOOLS

tool_icon NEBcloner®
Find products, protocols and tips for each step of your traditional cloning experiment.
tool_icon NEBioCalculator®
For help with scientific calculations and conversions

BROCHURES & TECHNICAL GUIDES

NEBNext_portal_literature_icon Molecular Cloning Technical Guide
Get help with product selection, protocols, tips for optimization and trouble-shooting.
NEBNext_portal_literature_icon Reagents & Tools for Molecular Cloning Brochure
Learn about NEB's recommended products for cloning.

 

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    Overview of Traditional Cloning

    Traditional Cloning refers to the generation of DNA fragments using restriction enzymes, and their subsequent assembly and transformation. The name is derived from the method’s history as the first widely-accepted cloning method. Learn more about the benefits and disadvantages of Traditional Cloning.