Procedures

From 2006.igem.org

Once this is completed it will be printed and available in the lab. It should include all the details so as to allow someone not familiar with a procedure to carry it out perfectly. Please add steps, comments etc. as you see fit.

Overnight Seeding:

• 1. Prepare desired volume of 1X LB all in one culture tube (calculate 1mL per seeding).
• 2. Add desired antibiotic (1uL per mL for Ampicilin).
• 3. Split the total volume by pipetting 1mL into additional culture tubes until each tube has 1mL.
• 4. Pick colonies and pipet up and down in LB to mix thoroughly.
• 5. Place in S-I overnight.

N.B.: Close tubes only partially to let air in.

Amplification Seeding:

• 1. Prepare desired volume of 1X LB + Antibiotic in culture tubes.
• 2. Pipet 100-200uL of overnight culture into the fresh LB.
• 3. Place in S-I until desired O.D. is reached.

Induction

Plating

Miniprep

Midiprep

Gel Electrophoresis

Gel Purification

Transformation

Ligation