Day 1: transformation
Preparation of LB agar plates with ampicillin 1/1000 and LB
Afternoon: E.Coli transformation –> Stratagene protocol: XL10-Gold Ultracompetent cells. Cat nº: 200314
PLASMIDS
A) MRGPRD genscript 10ug
B) GPR157 gescript 10ug
Reconstitution in 100ul sterile H2O
Day 2
Morning: Choose some good colonies and pick them with a tip.
Put it on a falcon tube of bacteria with 5 ml of LB + ampicillin 1/1000 -> release the tip inside the tube and leave it there.
Leave a few hours (6-8) at 37ºC stirring at 225-250 rpm (orbital shaker)
Prepare a Erlenmeyer of 150 mL LB medium: 3.75 g LB + 150 mL H2O. Autoclave and let cool. Add Ampicillin 1/1000
Day 3: maxiprep (kit)
- RESULTS
- MRGPRD (0.815ug/ul)
- GPR57 (0.891 ug/ul)
