DNA cloning—often used interchangeably with molecular cloning—is an umbrella concept that includes many different experimental techniques whose goal is to isolate and expand a specific fragment of DNA into a host organism to create a large number of identical copies (recombinant DNA).
DNA cloning was first described in the 1970s and has since evolved to become the cornerstone of biotechnology.
DNA cloning is comprised of multiple steps from generating recombinant DNA and vector constructs, to introducing them into host cells, to verifying the desired recombinant DNA final product. For this, DNA fragments are joined together and introduced into a vector which may include plasmids, attenuated viruses or bacteriophages, or larger constructs such as artificial chromosomes. Finally, the resulting recombinant DNA construct is transformed into host cells which use their own cellular machinery to replicate and express the construct.
This recombinant DNA technology is the foundation for a range of synthetic biology products like vaccines, biologics, enzymes, new crop varieties, microbial strains for bioremediation, research models, biodegradable materials, and biofuels, among others.
The basic cloning workflow involves these five steps:
There are numerous methods to insert a desired DNA fragment into a vector of choice. Each method has different applications and limitations—understanding which one is often a critical point in the experiment design step.
Synthetic, double-stranded DNA fragments are convenient alternatives to PCR products and can be used in cloning and gene assembly applications. These materials are flexible and customizable, allowing users to design and validate in silico while also adapting the fragments to specific requirements of the cloning method. This means that these synthetic DNA sequences are a convenient and faster option because they do not require amplifying or assembling off various templates that must already be available in the lab.
IDT offers three types of DNA fragments that are customizable and compatible with all cloning applications.
This guide discusses common DNA cloning techniques, important sequence design considerations, and tips for troubleshooting that may arise when performing a cloning experiment.
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