Week 2

July 18-

Colony grew, negative control didn’t

Transfomation- 50 µL DH5α, 2 µL DNA at 37 degrees Celsius overnight (all carb)

Glycerol stock- 350 µL of cells and 150 µL of 50% glycerol

Miniprep

Dissolving and diluting primers (10 pmol/µL)

PCR and gel electrophoresis and purification

Digestion for 1 hour and 30 minutes at 37 degrees Celsius

Gel Electrophoresis and Purification

Digestion and ligation

July 19-

Transformation- 50 µL DH5α, 2 µL DNA at 37 degrees Celsius overnight

July 20-

Incubate in 3 mL of LB and appropriate antibiotic overnight at 37 degrees Celsius and 220 rpm

Ligation of Photo and YFP (i2.5.4 and i2.5.1) for 10 minutes at room temperature

Transformation of Photo and YFP 50 µL DH5α, 2 µL DNA at 37 degrees Celsius overnight (all chlora)

Making competent cells- 20 tubes (250 µL each) RP437, 20 tubes (250 µL each) BW2513

July 21-

Glycerol stock- 150 µL of 50% glycerol and 350 µL of cells

Miniprep

Incubate in 3 mL LB and appropriate antibiotic overnight at 37 degrees Celsius and 220 rpm

Test digestion for 1 hour and 20 minutes a 37 degrees Celsius

- Test digestion for 1 hour and 20 minutes a 37 degrees Celsius

July 22-

Glycerol Stock- 150 µL of 50% glycerol and 350 µL of cells

Miniprep

Glycerol stock- 350 µL of cells abd 150 µL of 50% glycerol

Dissolving and Diluting primers

Test digestion and gel electrophoresis