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Team:HokkaidoU Japan/Notebook/plastic Week 2
From 2012.igem.org
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====Plasmid extraction==== | ====Plasmid extraction==== | ||
Extracted the plasmid. | Extracted the plasmid. | ||
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Revision as of 19:30, 26 September 2012
Contents |
July 9th
Transformation
Transformation of pGEM into JM109. This transformation is the bioplastic production test in JM109.
- Added 1 ul of plasmid DNA to 50 ul of thawed competent cells on ice.
- Incubated on ice for 30 min.
- Added 600 ul of LB.
- Prepared and Labeled two petri dishes with LBA.
- Plate 300 ul of the transformation onto LBA dish and spread.
- Added 900 ul of LB to 100 ul of the transformation and plated 300 ul of it onto LBA dish then spread.
- Incubated the plates at 37C for 15hrs 30 min.
July 10th
Incubation
- Single colony isolation.
- Pre-culture with 2xLB/A for 10~14hrs, 180 rpm, 30C .
- Cultured in plastic producing media for 48hrs, 180 rpm.
Polymer extract
Measure the weight of tube.
July 14th
Plasmid extraction
Extracted the plasmid.
