Modular Cloning Intro Jargon
Modular Cloning Intro
Why bother with modular cloning?
Modular cloning might not be necessary for every lab or project, but if you're planning to assemble more than two or three constructs, it can save you a significant amount of time and money.
Let’s consider a scenario: you want to clone three constructs, each with different combinations of promoters and signal peptides, but all sharing the same coding sequence and terminator. Using traditional cloning techniques, this could take anywhere from 2 to 4 weeks. Alternatively, you could order synthesized fragments, which might speed things up—but at a high price tag. Even for assembly when you factor in the number of primers and manhours needed to assemble all the combinations, the price can add up.
Now, imagine achieving the same outcome within that same 2–4 week timeframe, but with the added flexibility to generate every possible combination of your components. That’s the power of modular cloning.
With modular cloning, each component—promoters, signal peptides, coding sequences, terminators—is cloned into its own individual plasmid. This initial step takes just a few days to complete, including amplification, assembly, and sequencing. Once these parts are in place, you can mix and match them to rapidly build new constructs without starting from scratch each time.
This blog will walk you through:
- The modular cloning process
- Modular cloning platforms
- How to choose a platform for your lab
- Tips for organizing your parts and workflows
- and more
In addition to the basic vector construction terminology we established, there's a few more concepts you should know for modular cloning.
- Level 0/Base vector/Entry vector:
- Level 1 Acceptor:
This is the term for when multiple Level 0s (L0s) are assembled together. The backbone/vector that accepts them is called the acceptor of the cassette. Unless there's a whole transcriptional unit in a L0(in case of selection usually), Level 1s are usually single transcriptional units. Level 1 plasmids can be the final construct but aren't typically.
- Level 2 Acceptor:
This is the term for when multiple level 1 transcriptional units are assembled into a single plasmid.
These terms will be important to understanding modular cloning levels and organization of assemblies. Speaking of important concepts, let's talk about enzymes!
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