Team:HokkaidoU Japan/Notebook/plastic Week 2

Transformation

Transformation of pGEM into JM109. This transformation is the bioplastic production test in JM109.

1. Added 1 ul of plasmid DNA to 50 ul of thawed competent cells on ice.
2. Incubated on ice for 30 min.
3. Added 600 ul of LB.
4. Prepared and Labeled two petri dishes with LBA.
5. Plate 300 ul of the transformation onto LBA dish and spread.
6. Added 900 ul of LB to 100 ul of the transformation and plated 300 ul of it onto LBA dish then spread.
7. Incubated the plates at 37C for 15hrs 30 min.

Incubation

1. Single colony isolation.
2. Pre-culture with 2xLB/A for 10~14hrs, 180 rpm, 30C .
3. Cultured in plastic producing media for 48hrs, 180 rpm.

Polymer extract

Measure the weight of tube.

Plasmid extraction

Extracted the plasmid.