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 [math]\displaystyle{ CaCl_2 }[/math] 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.
