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Old 01-06-2013, 06:20 AM   #401
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Quote:
Originally Posted by jason.mundy

I haven't. I'm interested in doing it. Was a centrifuge needed to do this?


No centrifuge. The way I understand it is, after growing some yeast on a plate, pick a colony and transfer it to a slant filled with 2-3 ml of distilled sterile water and that's it. Without the agar. Close the lid and even leave it outside the fridge, will last for months, maybe years in the fridge. The reason I asked is, it kinda sounds too easy. Maybe I'm missing out on something....


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Old 01-10-2013, 08:28 AM   #402
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Starting my slants today.. hopefully


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Old 01-23-2013, 09:42 PM   #403
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So it's my first time.. ot a couple quick questions. I didn't end up slanting until this Sunday.. and I made a petri as well just to try different things. Is this what the petri dish is supposed to look like?

Also, I just realized today that I forgot to unscrew the caps a little.. they look pretty much the same as the dish with just a little liquidy streaking. I went ahead and cracked the caps a little with a puff of air and recovered them with tin foil in the jar.. will they still be ok or should I scrap them and try again next time or with scrapings from the dish?
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Old 01-23-2013, 09:58 PM   #404
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Quote:
Originally Posted by Roadliner View Post
So it's my first time.. ot a couple quick questions. I didn't end up slanting until this Sunday.. and I made a petri as well just to try different things. Is this what the petri dish is supposed to look like?

Also, I just realized today that I forgot to unscrew the caps a little.. they look pretty much the same as the dish with just a little liquidy streaking. I went ahead and cracked the caps a little with a puff of air and recovered them with tin foil in the jar.. will they still be ok or should I scrap them and try again next time or with scrapings from the dish?
Roadliner,
from what I can see in the photo, that is a lovely plate. But, I only see two, maybe three single cell colonies. That's what you are after. Collect about 10 of those and transfer by loupe to 2 ml steril wort and you will have purified (decontamiated) any yeast you may be using.
Concerning the cracking of the caps, I would say you are good, what with the egress of gas and covering with foil. If all was done with sanitary protocol, you should have no problems. Good luck in your yeast farming endeavers..

PS. Us Arkies gotta look after one another...
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Old 01-23-2013, 10:24 PM   #405
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Originally Posted by sudbuster View Post
Roadliner,
from what I can see in the photo, that is a lovely plate. But, I only see two, maybe three single cell colonies. That's what you are after. Collect about 10 of those and transfer by loupe to 2 ml steril wort and you will have purified (decontamiated) any yeast you may be using.
Concerning the cracking of the caps, I would say you are good, what with the egress of gas and covering with foil. If all was done with sanitary protocol, you should have no problems. Good luck in your yeast farming endeavers..

PS. Us Arkies gotta look after one another...
so all but the dots by themselves should not be used? lol guess I should have streaked a few more plates.. Next time I will
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Old 01-24-2013, 03:58 AM   #406
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Has anyone tried the "water immersion" technique from the "yeast" book? Page 198.
I have. That's all I keep now. My oldest resurrection to date is a sample that was stored something like 2 years and 4 months before I re-streaked it. The yeast woke right up, as if I had stored them just a week ago.

I do keep them in a fridge, just because the unused little shelf on the door is a convenient place for me to keep them. Every bit I've read, though, says I should expect the same result if I'd tossed them in a cigar box on desk.

http://www.homebrewtalk.com/f163/2-1...-yeast-351613/

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Old 01-24-2013, 02:59 PM   #407
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Does anyone know of any active slant trading posting going around here?
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Old 01-25-2013, 05:30 AM   #408
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there was one, but not much activity I guess.. I'd love to do some slant trading once I get my technique down.. but right now, I'm not even sure how to tell if my slants are ok to use or not...

Speaking of which.. how do I know if my slants are ok to use? i would hate to pour some nasty yeast into a batch and ruin it.
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Old 01-28-2013, 01:53 PM   #409
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My recipe for slant media is:

1.5% agar powder
7% DME
1% Wyeast yeast nutrient

That is by weight so if you have, say, 500mL of water you will add 7.5 grams of agar and 35 grams of DME along with 5 grams of the nutrient.

The agar powder def. works better than the stuff I got at the grocery store, which after a few months in the fridge started falling apart.
I'm not sure if this has been discussed in the pages after this post but I haven't gotten through all the posts yet. I mixed per the proportions above but my slants haven't set enough after 36 hours. My agar is from Now Nutrition and says 100% agar.


I boiled my DME and yeast nutrient for around 1 minute. Let cool for a few minutes then dumped in the agar powder and stirred. Then used a syringe to measure out for my vials. I pressure cooked as instructed, let cool 1 hour, tightened caps and taped (I am not looking forward to ripping tape off and retaping 36 vials! ).

The only thing I am suspicious of is my yeast nutrient. I used Fermax yeast nutrient. After boiling my wort, I got a strong ammonia smell. Perhaps Fermax is not the same concentration as the wyeast nutrient?

Edit: finally finished reading through all the pages. There appears to be a difference in the agar. Obviously the block stuff mentioned in the OP but also some flaked agar and even 2 different powder versions (a medical and the nutritional kind like I used). I also read something about getting the agar mix up to boiling temps to avoid separation while filling your vials. I did only check one vial so maybe others are more solid than the one I checked. So any input would be appreciated.

Regardless, I am curious about the Fermax issue I mentioned though.
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Old 01-29-2013, 05:28 AM   #410
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Quote:
Originally Posted by chuckjaxfl View Post
I have. That's all I keep now. My oldest resurrection to date is a sample that was stored something like 2 years and 4 months before I re-streaked it. The yeast woke right up, as if I had stored them just a week ago.

I do keep them in a fridge, just because the unused little shelf on the door is a convenient place for me to keep them. Every bit I've read, though, says I should expect the same result if I'd tossed them in a cigar box on desk.

http://www.homebrewtalk.com/f163/2-1...-yeast-351613/
I want to toss out there I do the oil immersion technique in YEAST now (started a couple weeks ago). I emailed white labs about it. I asked if the mineral oil used for butcher blocks was fine. And for the heck of it I inquired if 100% olive oil was ok. They never heard of using olive oil so did not recommend it. As far as the mineral oil, they just said mineral oil without being specific about 99.9% or 100%. I couldn't find any 100% mineral oil, but did find some 99.9% mineral oil for consumption (for constipation or something or other) at a rite aid or walgreens. The other .1% is vitamin E. They recommend it be sterilized before use, of course.

Anyhow, I poured some into a small mason jar and put it in the pressure cooker when sterilizing the vials and slanting media. It went in clear and came out with a white tint to it. I figure it was ok. After the slants grew appropriately, i put a few ml of oil in to cover the yeast and put it in the fridge. The only quirky thing is now I have to store the slants on an angle instead of upright so the oil stays on top of the growth area.


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