IGEM:Peking/2007/Count-Conjugation-Notebook/2007-7-14

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Contents

Result of The Potency Test of Competent Cells Made Yesterday

  • Plates with 100uL transformants: 5000~6000 (number of positive colonies).
  • Plates with 200uL transformants: 3000~4000.
  • Negative control: 0.
  • Note: it makes sense that the 200uL transformants grew less because I used 2 different tubes of competent cells. The less efficient one is weaker because it was treated with CaCl2 for a longer time(~ 1 hour).

Fetch the biobricks J01060, B0015 and R0040

  • Note: we should make a better design and plan so that there can be fewer cases where we found we need new biobricks.

Antibiotics Solution Preparation

  • Chloramphenicol: 34mg/mL, 10mL. solved in C2H5OH.
  • Ampicillin: 50mg/mL, 10mL.
  • Kanamycin: 50mg/mL, 10mL.
  • Stored at -20℃.
  • Note: All solution was filtered by sterile syringe filter, and was divided into 0.5mL tubes.

Transformation

  • Transformed R0040, J01060, B0015 into DH5alpha competent cells.
  • proT as positive control, empty cells as negative control.

Plans for tomorrow

  • Digestion Test of the extracted plasmid(The DNA dye has come! ^_^).
  • Miniprep of the rest biobricks plasmids.
  • Order a Fluorescence Intensity Test from the company.
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