Flasking questions

[greengarden]

Thanks, glad you turned up. Looks like an awesome method I'm already well on my way to using it

[The Orchid Boy]

zxyqu (Jarad) has a link to the reference sheet on the first page of this thread. I thought the video said you can't let dry seed sit in the bleach solution long at all. Just until most of the seed sinks and then rinse it... But you don't have to rinse the seed when using the peroxide. How do you know when the seed is sterilized? Thanks everyone.

[zxyqu]

Hydrogen Peroxide will take MUCH Longer (hours). 10% bleach solution and you're talking 10-15 minutes. 30 at max, at least with Phalaenopsis seeds... Much longer and you just have some stuff staring at you for months...

Seed sinking is a mere indication that all the trapped air on the surface of the seed has been displaced. It doesn't mean sterilized, but it assures you an air pocket on the seed didn't miss the sterilizing agent. That's a benefit of vigourously shaking, and adding a wetting agent (soap). You dont ever "know" the seed is sterilized for sure, that's why it's best to do several light sow mother flasks that get various times, or agents, of sterilization.

[The Orchid Boy]

Once I sow the seed, how long should it take to germinate? How can I know if it is no good? And if you don't sow lightly, you replate the protocorms right? What would happen if you don't sow lightly and don't replate or only replate once? I would like to avoid too much replating but will do it if necessary.

[zxyqu]

Lightly sowing mother flasks likely will not prevent having to do replates. If you get a ton of seed from a good pod, replates are just going to have to happen. However, if you only get a little seed, maybe you'll get lucky with only a couple hundred seed. Go ahead though and prepare to have to do replates at 6mo to a year, and possibly again, depending on how lightly you seed the replates.
Time to germinate varies, and I have no idea for Catts, but if it goes 3 months with no greening up of the seed, I'd imagine it's a bust.

[The Orchid Boy]

I'm just a little nervous about doing replates. I wish I could watch someone do them. If I don't sow lightly I replate a little while after the seeds become protocorms and every 1-2 months after that right? When replating do you remove all of the media from protocorms/flasklings, some of the media or none of the media?

[zxyqu]

Just take a clean scoop or sort, put them on new media and spread around. If you want to do this by yourself, you'll probably have to learn to do replates. You can do it in a glove box, or over sterile steam on a stove top. But it'll more than likely have to get done.

[The Orchid Boy]

Orchid media guide:http://www.phytotechlab.com/pdf/orchidmediaguide.pdf
I found out what the little numbers mean. The ^1 means media used at 1/2 strength. The ^2 means medium supplemented with up to 15% coconut water.

So does the media container have instructions for how to use at 1/2 strength and how much coconut water to add? The chart lower on the page says that P668 plus agar equals P658: ORCHID MAINTENANCE MEDIUM Contains Charcoal and Agar - Orchid Culture - P658 So could I use P658 for flasking and replating diluting and adding coconut water when the chart says instead of P668?

[zxyqu]

I'm not sure if you can use P658 at 1/2 strength as would be needed for seed sowing (as you'd dilute out the agar). Hence why I'd stick with 668 and agar. It's not that big of an issue once you are weighing them out. Plus, self addition of agar allows you to play around with gel density, which can have effect on seed growth.
Attached is exactly what my bottle of 668 says. 27.31 g/L is recommended, so I use 13.7 g/L for half strength, with agar, sugar, and water. Not sure what amount of agar is needed, as it'll depend on what you buy, but it should also provide the needed info on it's bottle (as well as on Phyto's website).

[The Orchid Boy]

So when using 13.7 g/L you use the same amount of water as you would use with 27.31 g/L but use the same amount of agar for both? You just use less of the powdered media? And that will make 1 liter of media?

Is g/L the same as grams? And it is weight right? That scale I have can measure in grams. To measure volume I have a glass meauring cup that goes from 50 mL and goes up by 25 mL at each line I might get some beaker from PhytoTech.

I found this method for water sterilization: http://missvickie.com/library/water-sterilization.htm Can I sterilize the media the exact same way?

Thanks so much for your time and patience answering these questions.