Team:HokkaidoU Japan/Notebook/plastic Week 2

From 2012.igem.org

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==July 9th==
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===July 9th===
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====Transformation====
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Transformation of pGEM into JM109. This transformation is the '''bioplastic production test in JM109'''.
Transformation of pGEM into JM109. This transformation is the '''bioplastic production test in JM109'''.
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#Added 1 ul of plasmid DNA to 50 ul of thawed competent cells on ice.
#Added 1 ul of plasmid DNA to 50 ul of thawed competent cells on ice.
#Incubated on ice for 30 min.
#Incubated on ice for 30 min.
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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.  
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#Incubated the plates at 37C for 15hrs 30 min.
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#Incubated the plates at 37C for 15 hrs 30 min.
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==July 10th==
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===July 10th===
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====Incubation====
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#Single colony isolation.
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#Pre-culture with 2xLB/A for 10~14 hrs, 180 rpm, 30C .
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#Cultured in plastic producing media for 48hrs, 180 rpm.
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====Polymer extract====
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Measure the weight of tube.
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===Cultivaton===
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#Single colony isolation
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#Pre-culture with 2xLB/A for 10~14hr, 180rpm, 30c
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#Cultured in plastic producing media for 48hr, 180rpm
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===polymer extract===
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#Measure the weight of tube.
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===July 14th===
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==July 13th==
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====Plasmid extraction====
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Extracted the plasmid.
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==July 14th==
 
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==miniprep==
 
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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.

  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 15 hrs 30 min.

July 10th

Incubation

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

Polymer extract

Measure the weight of tube.

July 14th

Plasmid extraction

Extracted the plasmid.


Retrieved from "http://2012.igem.org/Team:HokkaidoU_Japan/Notebook/plastic_Week_2"