Team:Queens Canada/Notebook/Protocols/Ligation

Ligation Storage and Labelling
- Label the product tube as “EC” and in accordance with the standard labelling format, as outlined at the front of your lab book and on Google Docs.
- Label the product plate as “EC (resistance)” and in accordance with the standard labelling format, as outlined at the front of your lab book and on Google Docs.

Materials
- Glass or plastic culture tubes
- Growth medium containing appropriate antibiotics
- Glass pipette tubes
- Parafilm

Procedure
1. Flame a glass pipette (or use a sterile plastic pipette without flaming), open the bottle of medium and flame the mouth.
2. Withdraw amount you need to fill your tubes (5ml per tube), flame the cap and recap the bottle as quickly as possible.
3. Remove the tube cap, flame the top of the culture tube, pipette in 5ml, flame the top of the tube and cap it.
4. Pick up one colony using a P20 pipette tip. Uncap the tube, flame the top, inject the tip into the tube.
5. Gently vortex the tube to ensure that the bacteria in the tip mixes into the media.
6. Incubate the tubes at 37⁰C overnight or until cells have reached the desired concentration. This should take between 12 and 16 hours.
7. When done, seal the transformed bacteria culture plate(s) that were used with Parafilm and store in 4⁰C fridge.