Setup and Staying Organized
Setting Up and Staying Organized with Modular Cloning
Modular cloning streamlines genetic assembly, but keeping your workflow efficient requires careful setup and organization. Following a structured approach will save time and ensure consistency.
Setting Up Your Modular Cloning Workflow
1. Choose a System and Stick with It
Selecting a single cloning system is crucial for long-term efficiency. While GreenGate and MultiGreen are interchangeable, most other systems are not. Switching between different platforms can lead to compatibility issues and extra troubleshooting.
2. Order the Necessary Materials
To begin cloning, you'll need to source the following:
Vectors: Most are available via Addgene, making ordering easy.
Reagents for ligation: Decide whether to include the enzyme in your order or purchase it separately.
Antibiotics: Essential for plasmid selection. If using CCDB, don’t forget chloramphenicol (Cm) for proper screening.
How to keep individual records and lab records
What to keep on your computer
Keeping individual records organized in a lab setting requires a consistent system—whether digital or physical. Use notebooks or electronic lab journals (ELNs) to record experimental details, observations, and protocols. Date all entries and include clear headings for easy reference. Maintain a backup of critical data, and categorize files logically with folders for raw data, results, and protocol changes.
To keep track of constructs and projects, organize your materials into clear categories:
Backbones – Storage for plasmid backbone sequences
L0 Library – Entry vectors for initial assembly
L1s & L2s – General-use plasmids for cloning and final assembly
Bridges/Linkers – Connectors for assembling multiple components
Primers – Designed for sequencing and verification
"Checking" & Raw Sequences – Reference files for validation
Each project should have a dedicated workspace where sequences, cloning steps, and progress can be tracked. A structured format could look like this:
Project #1:
✅ Relevant sequences & notes
✅ L0 plasmids needed for this project
✅ L1 plasmids to assemble
✅ Final plasmid for transformation
By maintaining clear project breakdowns, you ensure all elements of your cloning pipeline remain organized and easily accessible.
What to keep in the shared space
- plasmid map database
- Freeze stock strains and their locations
- eg:
- primer list and location
- eg:
Final Thoughts
Setting up a modular cloning workflow efficiently requires commitment to a system, proper reagent ordering, and organized project tracking. Whether working with MoClo, GoldenBraid, or MultiGreen, using structured tools ensures smoother cloning and faster progress.
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