Team:HokkaidoU Japan/Notebook/overall protocols

From 2012.igem.org

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==Transformation==
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#Add 1~2 ul of DNA to 50 ul of thawed competent cells on ice.
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#Incubate on ice for 30 min.
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#Add 600 ul of LB.
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#Incubate the cells for 2 hours at 37C if added DNA plasmid have antibiotic resistance other than ampicillin.
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#Prepare and Label two plastic plates with LB and appropriate antibiotic.
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#Plate 300 ul of the culture onto first dish and spread.
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#Add 900 ul of LB to 100 ul of the culture and plate 300 ul of it onto second dish and spread.
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#Incubate the plates at 37C for 16~20 hours.
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Revision as of 09:54, 10 August 2012

Transformation

  1. Add 1~2 ul of DNA to 50 ul of thawed competent cells on ice.
  2. Incubate on ice for 30 min.
  3. Add 600 ul of LB.
  4. Incubate the cells for 2 hours at 37C if added DNA plasmid have antibiotic resistance other than ampicillin.
  5. Prepare and Label two plastic plates with LB and appropriate antibiotic.
  6. Plate 300 ul of the culture onto first dish and spread.
  7. Add 900 ul of LB to 100 ul of the culture and plate 300 ul of it onto second dish and spread.
  8. Incubate the plates at 37C for 16~20 hours.



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