TC medium for drosera OK?

[klasac]

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.
Some chemicals:

TC :

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?
THANX

 

[jimscott]

Egads that's complicated! And to think we have pyridoxine, kobalamin, mannitol, isopropanol, and hydrogen peroxide in our lab... along with WFI water.

 

[klasac]

Well we have this in our lab. Those are quite common chemicals. And we are not even a biochemistry lab. We are organic chemistry lab. So I thought someone from the forum might have an opinion on whether I have done it right and try to answer my questions. I see tissue culturing is not as common (or popular?) in amateur circles as I thought it might be. Well no problem...time will show the outcome of my debut experiment... Thanx anyways.

 

[CPlantaholic]

It's not all that popular yet, but there are still plenty of people in the cp community that use tissue culture. The only difference is that instead of using leaf cuttings (which are a bit harder to use in tc), seeds or root cuttings are more commonly used. The reason for this, is that it is harder to sterilize the leaves without damaging them, compared to roots, which are more durable and thick. The leaves look pretty good to me, so I'm sure you'll have luck with them.

 

[klasac]

Thanx plantaholic. I know there is a thin line between disinfection and killing of the tender leaf tissue. That is my main concern as well. I will report later which it was