Team:Montreal/Seeding

From 2008.igem.org

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Protocol

Seeding is a method for amplifying a single colony of genetically identical bacteria.

1. Pipet 5.0 mL of LB broth into two 10 mL culture tube, under sterile conditions.

2. Add the applicable antibiotic (amp+, 1 µL/mL LB; kan+, 5 µL/mL)

3. Identify an area on the transformation plate with single colonies. "Pick" one colony by picking it up with a sterile P200 pipet tip, and eject the tip into the media of the culture tube.

4. Leave one tube with only LB and antibiotic as a control

5. Fix the caps on the tubes loosely, to allow air circulation and subsequent bacterial growth.

6. Place the tubes in a shaking incubator at 37oC for 16-20 hours.