We have collated and stored a collection of plasmids and yeast strains that represent useful tools used in the development of our TR&D projects as well as being instrumental for our Driving Biological Projects and Collaboration and Service Projects called the Clone database. This resource is open to the biological community. And we are constantly expanding this resource to better serve the needs of the scientific community. Importantly:
- Many of the plasmids in this resource encode bacterially-expressible nanobodies against various antigens. If you are interested in any of these, we encourage you to engage with us as we likely have additional information concerning the best ways to express and purify these nanobodies and biophysical data on their characteristics.
- If you do not see the exact construct you need, but there exist related ones, contact us as it is possible we have another unlisted version that may better suit your needs, or we can easily construct a version suitable for your needs.
NCDIR deposited a set of 34 GFP and mCherry tagged nanobodies to Addgene. These nanobodies will become part of a special affinity purifications section of Addgene.
https://www.addgene.org/Michael_Rout/
An excellent primer on plasmids and their uses is available at: https://www.addgene.org/mol-bio-reference/
Storage
Plasmids
You may receive our plasmids in the form of DNA on filter paper or as liquid aliquots in tubes. Approximately 0.5 µg of DNA will be sent, and we recommend amplification before use.
DNA on filter paper in tubes can be stored at 4°C. To recover the plasmid on filter paper:
- Use a clean razor blade to cut out the marked circle containing plasmid DNA.
- Immerse the circle in 30µL of TE and pipette to mix.
- After waiting for at least 10 minutes, use 2µL to transform competent bacteria.
Liquid DNA suspensions received in tubes can be stored at -20°C. We recommend transforming the plasmid into E. coli and performing minipreps to amplify the plasmid. For long term storage, keep the purified plasmid DNA at -20°C. You may additionally choose to make a bacterial glycerol stock (final glycerol concentration can be 15 to 25% v/v) to be stored at -80°C.
Yeast stains
You may receive our yeast strains grown on an agar plate.
Yeast strains can be maintained as colonies on agar plates at 4°C. They need to be restreaked every 2 to 4 weeks. For long term storage, freeze the strain in 15 to 20% (v/v) glycerol and store at -80°C.
Using Our Strain or Plasmid
The paper originally citing the reagent will also have methods detailing its use.
If you have any other questions, contact the corresponding author of the paper or NCDIR administrator: Admin@NCDIR.
Antibody Verification
We have verified each reagent at the time of deposition. However, if troubleshooting is needed, we can recharacterize the reagent upon request.
Citations in Future Publications
Please acknowledge the Principal Investigator, cite the original article listed on the clone database, and include appropriate grant numbers listed in the original article.
Contact NCDIR
If you have questions or would like to speak with an NCDIR researcher, please contact us at Admin@NCDIR