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

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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 Failed to parse (MathML with SVG or PNG fallback (recommended for modern browsers and accessibility tools): Invalid response ("Math extension cannot connect to Restbase.") from server "https://api.formulasearchengine.com/v1/":): {\displaystyle CaCl_2} 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.