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Home Brew Forums > Home Brewing Beer > Lambic & Wild Brewing > Howto: Capture Wild Yeast
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Old 10-15-2012, 12:56 PM   #801
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Ok after about 1.5 weeks of capturing some local bugs I have a fairly thick layer on the bottom of my mason jar pretty comprable to washing yeast. The smell is not at all unpleasant it almost smells like plain yogurt very lactic and somewhat sweet smelling initially somewhat honey like though there was only Dme and water. What I found interesting was that there were no bubbles like in a regular fermentation, does this mean I didn't catch any yeast that all i got was bacteria ? should I put it back outside ?

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Old 10-15-2012, 07:30 PM   #802
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As long as you have something, I'd consider running a teaspoon of the sediment through another starter. It should behave similar to a normal starter as in it should turn cloudy and then start to settle within three or four days. If you behavior takes weeks or if it smells wretched on the second starter, then I would suspect you don't have what your looking for.

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Old 10-16-2012, 06:59 AM   #803
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Ok after about 1.5 weeks of capturing some local bugs I have a fairly thick layer on the bottom of my mason jar pretty comprable to washing yeast. The smell is not at all unpleasant it almost smells like plain yogurt very lactic and somewhat sweet smelling initially somewhat honey like though there was only Dme and water. What I found interesting was that there were no bubbles like in a regular fermentation, does this mean I didn't catch any yeast that all i got was bacteria ? should I put it back outside ?
i have exactly the same thing. mine never bubbled or got foamy, then started to smell cheezy, now after 3 weeks i have convinced myself it smells sour/spicy. gf thinks it smells like cold vomit. i keep flipping between, "i've done something really disgusting here" and "total breakthrough!". i'm going to make a very small starter and add a little bit. all suggestions will be considered! i will also streak some out on plates and have a good hard stare at them
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Old 10-17-2012, 01:42 PM   #804
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I can't wait to start doing this again. I just moved out of home, and now I have to go and get some DME...

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Old 10-18-2012, 05:43 PM   #805
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After stepping it up in a second starter, my homegrown guys are going to town in a one gallon test batch brewed with my backyard Chinook hops. The scent on the yeast starter was lemony/pineapple. We'll see how this goes...

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Old 10-18-2012, 10:53 PM   #806
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This thread is awesome. I have an experiment on going and will post photos soon, but I had a question.
I have a few strains of wild yeast that metabolize bromocresol green - so they are white/opaque when grown on BCG, yet they are sensitive to cyclohexamide at 10-20 mg/L.
From what I've read, Brett should be resistant to cyclohexamide. Any ideas to what these yeast might be. They are clearly budding yeast.
Thanks.

Doing some more searching, it seems possible that they might be Candida? That would suck as all my isolates have the same phenotype. One more edit. I don't think many Candida can tolerate cycloheximide.

Another update. I actually got one clone after 3 days at 30 degrees C that grew on cyclohexamide (last picture). It's looks no different than (colony morphology or cells) the cyc negative yeast.
I grew small cultures (10 mL in DME with a small bit of yeast extract) and neither of the cultures have much flavor at all? Do I need larger/longer grows? Yes, I might need to grow the yeast on cycloheximide for 7-10 days.

Ok, here are some photos of two wild yeast clones isolated from different sources.
What do you guys think? I think they might be Brett.

rsf3jpg.jpg
Clone 1
rsf3a-jpg.jpg
Clone 1
jbcycajpg.jpg
cyclohexamide resistant Clone 2

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Old 10-21-2012, 10:22 AM   #807
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can you send samples to a yeast lab for genotyping? i am very familiar with plant genotyping techniques and molecular biology in general, it's relatively simple to amplify a bit of ribosomal RNA genes or microsatellite DNA and sequence it, so i would imagine that a well equipped lab that already knows which bits to sequence could knock them out easily and give you a genus?? but i know nothing about yeast genotyping. if you are buying in bulk the total cost of reagents+materials to sequence a gene from one colony on a plate is probably around 20 dollars with the economy of scale (and if you happen to own a dye-terminator capillary sequencer!) and can be done from the amount of cells on the end of a toothpick stuck into the colony.
my point is it might not cost that much if there is a willing lab locally

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Old 10-21-2012, 02:16 PM   #808
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That really brings up a great point.... Is there a lab that anyone knows of to perform this identification. As well to do this service for under $1,000's of dollars. I would love to identify what I have caught as I will be slanting and isolating colonies.

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Old 10-21-2012, 03:16 PM   #809
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That really brings up a great point.... Is there a lab that anyone knows of to perform this identification. As well to do this service for under $1,000's of dollars. I would love to identify what I have caught as I will be slanting and isolating colonies.
Siebel does it for 200 dollar or so. All they do is just a simple PCR of a specific part of the genome. I do not have the info here, but will post it next week. I tried the ITS parts myself as well, but those are hard to sequence because of the repeats. They use a different sequence.

I made a protocol where you can just use some yeast and PCR on them without doing DNA isolations. Yeast is a pain to lyse, but it worked. Then sequencing of the PCRed DNA is easy. A simple blast will give you your results.
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Old 10-21-2012, 04:11 PM   #810
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Siebel does it for 200 dollar or so. All they do is just a simple PCR of a specific part of the genome. I do not have the info here, but will post it next week. I tried the ITS parts myself as well, but those are hard to sequence because of the repeats. They use a different sequence.

I made a protocol where you can just use some yeast and PCR on them without doing DNA isolations. Yeast is a pain to lyse, but it worked. Then sequencing of the PCRed DNA is easy. A simple blast will give you your results.
That would be great if you post that info it would be much appreciated.
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