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

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 $$CaCl_2$$ 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.