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Team:HokkaidoU Japan/Notebook/plastic Week 2
From 2012.igem.org
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| - | ==July 9th== | + | ===July 9th=== |
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====Transformation==== | ====Transformation==== | ||
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#Plate 300 ul of the transformation onto LBA dish and spread. | #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. | #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 | + | #Incubated the plates at 37C for 15 hrs 30 min. |
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<div class="hokkaidou-notebook-daily"> | <div class="hokkaidou-notebook-daily"> | ||
| - | ==July 10th== | + | |
| + | ===July 10th=== | ||
<div class="hokkaidou-section"> | <div class="hokkaidou-section"> | ||
====Incubation==== | ====Incubation==== | ||
#Single colony isolation. | #Single colony isolation. | ||
| - | #Pre-culture with 2xLB/A for 10~ | + | #Pre-culture with 2xLB/A for 10~14 hrs, 180 rpm, 30C . |
#Cultured in plastic producing media for 48hrs, 180 rpm. | #Cultured in plastic producing media for 48hrs, 180 rpm. | ||
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====Polymer extract==== | ====Polymer extract==== | ||
Measure the weight of tube. | Measure the weight of tube. | ||
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| - | ==July 14th== | + | ===July 14th=== |
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====Plasmid extraction==== | ====Plasmid extraction==== | ||
Latest revision as of 03:55, 27 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 15 hrs 30 min.
July 10th
Incubation
- Single colony isolation.
- Pre-culture with 2xLB/A for 10~14 hrs, 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.
