pre sterlized agar?

[flhiker]

My first attempt at flasking didn't go so well. I grew mold of all colors (not my goal). So while looking for agar online I found someone selling this kit. I realize this is more for experiments but what do you think? The media is pre autoclaved and all you have to do is microwave the solution and pour it in a petri dish.
Nutrient Agar Science Fair Plate Pouring Kit - eBay (item 140303953169 end time Mar-04-09 08:07:46 PST)

Any Thoughts. I know I'm lazy

[Gin]

Hi , It does not sound like it has the proper nutrient additions for orchid seeds also you would still be stuck re plating would be back to square one .
Try it on your own again until it works . Where I had trouble was with the re plating grew some wild stuff the seedlings were not in it yet.Re plate flasks had sat for almost a month before going bad . It was my fault the way I vented them . Gin

[Royal]

Hey Dave,

I would tend to think that pouring sterile media into sterile vessels would be harder to keep out contamination than just autoclaving closed vessels. You'd have to pour them up in a sterile environment, and unless you have a nice roomy flow hood I don't see this option working too well. I wouldn't even consider it for my conditions (kitchen).

You want to eliminate as many variables as possible to pin down where your contamination is coming from. Most of the time it comes from us opening the flasks and transferring to and from them (incindental). But you can't know this for sure unless you are positive they were sterile before you sow seed or replate.

So, make up a batch of flasks and let them sit for 7-10 days. If they contaminate, then you either have issues with the closures (lids or vents) or you're not achieving high enough temps for a long enough time in the p-cooker.

If they look good after a couple of weeks, your in the clear. Next, when you do your next batch, bring an extra flask into your workspace. Open this extra one up to expose it to the glovebox environment. Once you finish sowing/replating, close up the extra flask. If this one contaminates then your workspace isn't "clean" enough.

If the extra one looks good, but the seeded flask contaminates, then you may have issues with tools, inadequate seed disinfection, or endogenous contamination of the seeds themselves. I hope this makes sense.

[flhiker]

Thanks Gin and Royal. Yes Royal it makes perfect sense. Use the extra flask as a control. To narrow down the source of the contamination. When sterilizing the seeds how long do you leave them in the bleach solution? And would you sterilize green unopened pods?

[Royal]

I haven't sown from green capsules that much, but yeah, I do sterilize the capsule first. I wash with dish soap and scrub gently with a soft toothbrush - just my fingers if it's fragile. Then I give it a soak in bleach. You can go pretty strong for this treatment, maybe 1/4 bleach? For me, green capsules don't go so well so maybe someone else can help there.

For dry seed I soak for 5 to 10 minutes, usually about 7. I just use commercial strength (6%) and dilute it to 1:10.

What are you working in? That was a tough problem for me. I've worked out some kinks, but I have a long way to go still. At least if you can lick the contamination issue you can focus on more productive things, like germinating those little buggers (and getting out in the wild and snapping more of those photos we've been waiting all winter for ).

[flhiker]

Originally Posted by RoyalOrchids

I haven't sown from green capsules that much, but yeah, I do sterilize the capsule first. For me, green capsules don't go so well.

For dry seed I soak for 5 to 10 minutes, usually about 7. I just use commercial strength (6%) and dilute it to 1:10.

What are you working in? That was a tough problem for me. I've worked out some kinks, but I have a long way to go still. At least if you can lick the contamination issue you can focus on more productive things, like germinating those little buggers (and getting out in the wild and snapping more of those photos we've been waiting all winter for ).

I have a glove box in the works (melamine, plexiglass, rubber gloves). other then that I'm like you I will be in the kitchen because that's where the stove is. As for the Pictures, I've been out but I thought everyone was getting tired of them, because they are mostly the same type of plants. Of course picture's really do not do the Strand justice. So I can see why others would be bored with them. I will upload some new finds in my gallery.

[Gin]

Bored with your pictures (never) keep them comming

I also use my kitchen , the capsule is scrubbed in bleach water with a soft tooth brush , agar and distilled water is brought to a boil on top of the stove then transferred to the containers , containers are then put in the pressure cooker 15 lb for 15 minutes. Gin