9:00-9:50: ML1 Insertion activation line 61~121 preparation: Tape the number tag.
9:50-10:50: Make ecotype stocks (3 and 4).
10:50-11:00: Set up plate sterilization (as described before).
11:00-11:40: Wash and rinse plates.
11:40-12:00: Wash and dry petri dishes.
12:00-13:40: Inoculate plants with 0.1A ES524 using syringe.
14:20-17:30: Make ecotype 3&4, ML1 Insertion 61~121 stocks in 96-well plates:
ecotype: Add 200 micro litre MS+MES media to two plates with ecotype seeds.
ML1: 1) Add 250 micro litre media to each tube with seeds;
2) Transfer 50 micro litre solution from each tube to the well of 96-well plates (four replica prepared);
3) Add 150 micro litre media to each well.
4) Add 200 micro litre media to the outer wells without seeds.
Stick aroud the plates and freeze in fridge.
17:30-18:10: Transfer leaf tissue from inoculated plants to petri dish to obtain bacteria growth curve.