This is composition of tissue culture agar medium I will attempt to propagate droseras in:
MACRO-elements: CaCl2(100mg), KH2PO4(60mg), KNO3(630mg), MgSO4(60mg), NH4NO3(550mg)
MICRO-elements: CoCl2.2H2O (0,01mg), FeSO4.7H2O (10,0mg), CuSO4.5H2O (0,01mg), H3BO3 (3,0mg), KI (0,40mg), MnCl2 (10,0mg),Na2MoO4.2H2O (0,1mg), ZnSO4.7H2O (4,5mg)
VITAMINS:thiamin hydrochloride,B1 (0,1mg), riboflavin,B2 (0,09mg),nikotinamide=niacin,B3 (0,95mg), pyridoxine,B6 (0,13mg), biotin,B7 (8mikrog),kobalamin,B12 (0,05mikrog),folic acid,Bk (0,01mg), panthotenic acid,coA (0,352mg), ascorbic acid,C (3,17mg)
FYTOHORMONES: NAA (0,05mg), IBA (0,09mg), IAA (0,1mg)
MEDIUM AND NUTRITION: agar (7,0g), maltodextrose (7,0g), pentaerytritol (3,0g), D-mannitol (10,0g), sucrose (3,0g)
DISINFECTION growing vessels, glove box and utencils: 1.Ca(ClO)2 2. isopropanole 3. 20minutes UV radiation 4. autoclave (20 mins at 120C) 4. (twizzlers-flame and alcohol)
DISINFECTION of explantates: 1. isopropanole (70%, 2 mins) 2. I2/KI/H2O (20mins) 3.hydrogen peroxide (5%, 5 mins + atomized oxygen) 4.triple rinse in disinfected RO water.
I have never done this before and I am not a biochemist so I have some questions for someone who is more experienced than me:
1. how long can store ready solution of agar medium of above listed composition in a fridge in a brown-glass bottle?
2. do the vessels with tissues need to be air-proofly confined in order to prevent infection of the inside by bacteria/fungi cultures?
3. If contaminated,how long after laboration will fungus/bacteria appear?
4. What is usual average time for firts observation of a new growth?