pUC19 Competence Results
Transformation efficiencies for pUC19 in ultra competent and normal competent cells. Needs improvement. Protocol updated with suggestions for getting better efficiency.
Ultra-competent: 2.1 x 108 transformants/ug DNA
Normal competent: 4.1 x 106 transformants/ug DNA
Transformation
Transformed Golden Gate products for Lyn+stop, Lyn-mKate, Lyn-TEVp, Syk-eYFP, Syk-TEVp into ultra competent cells. Plated 200uL of each at 5pm. Removed plates from 37C incubator at 9am.
Liquid culture
Started 5 liquid cultures each (5mL LB + 5uL kanamycin) for CD79A+stop, CD79A-TCS-Gal4VP16, Syk+stop. No blue colonies were observed on the plates and there was no positive control of just GGDonr backbone, so the Golden Gates will have to be redone. In the mean time, continue as though the colonies are correct. Placed in shaking incubator (37C) at 4:50pm. Removed from incubator to refrigerator at 9am.