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Schlenkerla

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I just started yeast culturing and I got to say I really like it. Two weeks ago I made slants and last week innoculated the slants and watched the yeast grow from what appeared to be nothing to white colonies of yeast.

You know what impresses me the most, I also innoculated about 50ml (~1/4 cup) of wort with a wire loop just dipped in a wyeast smack pack. The mini-starter was foamy in 2 days, then 2 days later I upped to 250ml, thursday I bumped it to 1000ml.

Now the starter is really kicking. All with just a dip in the smack pack!!!



The reason I really like doing this, its like brewing all week as you step up the starters. Now I'm all set for tonights brew session. A Cream Stout w/ wyeast 1028 London Ale Yeast.

Here's how the slants are doing. (Both have fluid in them which happens to be internal condensate from the pressure cooking. )

Slant #1



Slant #2



I'll be set on yeast for years to come... Wyeast 3638 Bavarian Wheat is next :ban:
 
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Schlenkerla

Schlenkerla

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Ok - None of you guys are doing this or considered it ever???

I'm just tired of paying $6-8 for yeast each time. Not too mention that I can have a bank of a dozen yeasts.
 

Iordz

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I culture, so I'm interested. I think it's fun, plus you can save money. But my favorite reason is having some rare strains (French saison, Pacman, etc) all year. I think you poured the agar-wort mix while it was too hot. Try pouring the mixture while it is around 100F next time, you will get a lot less condensation.
Looks good!
 

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Schlenkerla said:
Ok - None of you guys doing this or considered it ever???

I'm just tired of paying $6-8 for yeast each time.
I haven't plated/slanted any yeast for homebrewing because when I've worked with yeast before (in a research lab) they didn't do so well after about a month if they were kept on solid media (at -28C). At -80C it will last forever in liquid media, but I don't think I'll be stepping up to that kind of freezer anytime soon. For now I'm just using expensive yeast in liquid media for a few months in the freezer.
 

Iordz

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I would highly recomend storing slants in a refridgerator, never a freezer because it will kill the yeast. I store mine in the fridge and "re-slant" them every 6 months, to ensure viability. I have heard that slants can be good for over a year if stored properly.
 
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Schlenkerla

Schlenkerla

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Iordz said:
I think you poured the agar-wort mix while it was too hot. Try pouring the mixture while it is around 100F next time, you will get a lot less condensation.
Looks good!
Thanks!

I was wondering why this was like that. I boiled the DME & Agar then turkey basted it to the test tube. After that I capped them, then pressure cooked the suckers. I cooled them on the rack at a 45 degree angle.

I heard you can keep the lids loose too, so they dry better.
 
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Schlenkerla

Schlenkerla

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Iordz said:
I would highly recomend storing slants in a refridgerator, never a freezer because it will kill the yeast. I store mine in the fridge and "re-slant" them every 6 months, to ensure viability. I have heard that slants can be good for over a year if stored properly.

I read on a few sites that storing in sterile water works pretty good. I'll be trying the Aussie Craftbrewer method.

http://www.brewery.org/brewery/library/SterileDW1096.html

http://oz.craftbrewer.org/Library/Methods/Lacey/YeastHand.shtml
 

Iordz

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You know, I tried the sterile distilled water and it didn't work well for me, so I moved back to slants. The problem was the yeast would go dormant, which is what you want, but then I couldn't really get enough to make a healthy starter. I like the fact that the yeast grows on the slant media, so there's more yeast whereas in the water the just go dormant.
Try it out and let me know if it works out for you.
 

CodeRage

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I was doin slants a while ago but they're too much of a pia to maintain. I want to freeze some but for some reason no one carries glycerine. Slants and petri dishes seem to be a good way to determine if the yeast are still good, plus you can isolate a few single colonies if you really wanted to.
 

Kubed

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I admit it Schlenkerla, after seeing a picture of your starter I was motivated to go start one of my own.

 

fratermus

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Schlenkerla said:
Ok - None of you guys are doing this or considered it ever???

I'm just tired of paying $6-8 for yeast each time. Not too mention that I can have a bank of a dozen yeasts.
Following along with great interest, because science gear is good geeky fun (beakers? loops? petri dishes? bunsen burners? I'm in!) and because it seems wasteful to underutilize a commercial yeast culture.
 

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Virtuous said:
I was doin slants a while ago but they're too much of a pia to maintain. I want to freeze some but for some reason no one carries glycerine. Slants and petri dishes seem to be a good way to determine if the yeast are still good, plus you can isolate a few single colonies if you really wanted to.
you should be able to find it at any pharmacy.

i love culturing yeast as well, the main part that drives me nuts is boiling up starter medium every few days during the week. i think i might start canning it so i can just do a bunch at once and pop em open as i need them.
 

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Kubed said:
I haven't plated/slanted any yeast for homebrewing because when I've worked with yeast before (in a research lab) they didn't do so well after about a month if they were kept on solid media (at -28C). At -80C it will last forever in liquid media, but I don't think I'll be stepping up to that kind of freezer anytime soon. For now I'm just using expensive yeast in liquid media for a few months in the freezer.
That's what I do, since I have a -80 where I work. Once I make my starter of a new yeast strain, I pitch about 3/4 of it, then build the starter back up to about 2 liters. Then I bring it into lab, spin down the yeast, then freeze in a 35% spent wort, 15% glycerol, 50% yeast solution.
 
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Schlenkerla

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scottfro said:
you should be able to find it at any pharmacy.

i love culturing yeast as well, the main part that drives me nuts is boiling up starter medium every few days during the week. i think i might start canning it so i can just do a bunch at once and pop em open as i need them.
I know what you mean. I thought of this too. To make it simple I've gone metric. I use the 10:1 rule. 10ml water : 1 gram DME. Gets you 1.040

I also boil in the flasks. I hate the part of waiting cool to do anything.

At lunch today I bought some tiny heinz ketch-up bottles.



I'll put 50ml of cold water in each then 5 grams of DME. Throw them in the pressure cooker w/ the the lid on and a splash of vinegar and boil the suckers for 20 minutes.

This way all I have to do grab one of these and clean the outside to near sterile. Then drop a loop of yeast in, cap it, and wait. I'll burp'em once a day or leave the cap loose.

The thing w/ the boiling is that it gets the flasks to safe level of cleanliness w/o much effort. The smallest ones are the hardest to do since you can't fit them on a gas burner very easily.

Question: I was told that erlenmeyer flasks are not supposed to break from thermal shock if you crash cool them. I haven't tested this since I don't want to learn the hard way.

Do any of you do this?
 

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Schlenkerla said:
Question: I was told that erlenmeyer flasks are not supposed to break from thermal shock if you crash cool them. I haven't tested this since I don't want to learn the hard way.

Do any of you do this?

Only high-quality borosilicate erlenmeyer flasks (pyrex #7800 glass) are resistive to thermal shock. Even then, I've still had flasks break on me (enough that I'm still always nervous when I do it).
 

Kubed

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mrkristofo said:
Only high-quality borosilicate erlenmeyer flasks (pyrex #7800 glass) are resistive to thermal shock. Even then, I've still had flasks break on me (enough that I'm still always nervous when I do it).
I'll 2nd that! I've broken more glassware with safety ratings than I can count. When I cool a hot flask I put it in an inch or so of cool (not cold) water, give it a few min, then start adding more water and ice to the bath.
 
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Schlenkerla

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Kubed said:
I'll 2nd that! I've broken more glassware with safety ratings than I can count. When I cool a hot flask I put it in an inch or so of cool (not cold) water, give it a few min, then start adding more water and ice to the bath.
OK - I can drop a hot flask on warm water, then repeat this once most of the heat is gone. Basically lessen the shock!!
 

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I hit pyrex erlenmeyers at full boil with very cold tap water in the lab all the time. 500ml size and I have yet to lose one. I also do this with the 1000ml flasks I have.

But I don't go that extreme with my 5000ml from morebeer. I use warm water to bring it down a little slower.
 

Kubed

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mr x hit the key point on this, and that's the volume of hot fluid you're dealing with. If you have 100ml in a 1000ml flask, it will cool quick and you prob won't have a problem. However when I have 800-900ml in a 1000ml, and it's my hand that's placing it in an ice bath, I'll take a few extra min to cool it down
 

fratermus

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Iordz said:
I think you poured the agar-wort mix while it was too hot. Try pouring the mixture while it is around 100F next time, you will get a lot less condensation.
Looks good!
First time slanter here.

I am cooking up the starter, piping into the slant tubes, capping loosely, and putting them at the 45deg angle in the pressure cooker (tube stand wedges in perfectly), and doing the 15lbs/15mins thing. I let it cool overnight then open up and seat the stoppers more securely.

I was doing it this way to get the least amount of infection exposure, but I did get condensation in the tubes. Would it help if I just kept the stoppers loose in the cooker and capped the beakers when cool? Would this let more steam escape and therefore produce less condensate?

My access to relatively dustfree areas is limited, so I really don't want to get into free-pouring sterilized wort into sterilized tubes if I don't have to.

What are my options here?
 

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Schlenkerla said:
Ok - None of you guys are doing this or considered it ever???

I'm just tired of paying $6-8 for yeast each time. Not too mention that I can have a bank of a dozen yeasts.
Yes I am culturing too. You will also notice that when the proper quantity is pitched the fermentation takes right off and is almost violent unless lower temperatures are used for fermentation which is a good thing. Be sure to label well as after you get 20 tubes, it get's confusing. :)
 

doublegun

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we cultivate here as well. More yeast strains than vegetables in our fridge. Really appeases the inner mad scientist.
 

fratermus

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WBC said:
Be sure to label well as after you get 20 tubes, it get's confusing. :)
Luckily the tubes I got off eBay (100 of them for $10!) have a silkscreen area to write on. I used a grease pencil on them to mark a number on them.

BTW, here's a pic of my tube with heavy condensation. Suggestions welcomed.

These are pretty big tubes and the #2 stoppers are really too small. I've got a pound of #3 stoppers on the way from Basic Science Supplies. I've bought from them a few times and am happy with price, shipping (10% of order, minimum $7), etc. Stuff I can't get there I buy off eBay or one of the IHBS.
 

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Is there a wiki for this method anywhere? I would love to get a step by step as far as the making of the agar for the tubes. Right now I currently just clean my yeast but this seems to be much more efficient.
 

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I culture yeast. I have a couple of strains that are hard to get and that I use quite often. That makes it worth it. I also have swapped yeast with other mad scientists and that is great fun. I too have noticed that yeast that I propogate up from a slant is very active and much more healthy compared to the Wyeast or WL.

To solve your condensation problem monitor the temp of your agar medium. I've noticed that to completly melt my agar I have to bring it up to boiling, but it will stay melted to about 120F. So, I process my agar in the pressure cooker, pull it out and then let it cool with a temp probe stuck under the vessel. When the agar medium cools below 150F I pour my plates and slants. I still will end up with condensate, but not nearly as much.

Another thing you can do to remove condensate is to remove the tops and lay your plates down on paper towel upside down and propped up on the neighboring plate. This will allow the condensate to evaporate. I put mine in a my clean box in this manner for 8 hours and then they are fine. (Note if you are not using a clean box or other clean environment, it is easy and cheep to make a clean box out of a clear plastic tub.)
 

fratermus

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wedge421 said:
Where do you guys get the agar from?
Asian food store. 25g for less than $1. 39 for the packets I picked up most recently. at a few grams for a batch, it goes a long way.

Also picked up a cheap 1/10th gram scale off eBay for something rediculous like $15 shipped. Works fine.
 

fratermus

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fratermus said:
Would it help if I just kept the stoppers loose in the cooker and capped the beakers when cool? Would this let more steam escape and therefore produce less condensate?
Tried this; didn't work. Might have even had more condensation.

Since I am forced to work in a dusty environment, I will try stuffing cotton in the tops of the tubes, pressure cooking them, and letting them sit for a week or so and see if it evaporates. If it does (and stays uninfected), I'll cook a set of stoppers for them, remove the cotton, innoculate, and cap.
 

fratermus

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Brickhouse said:
To solve your condensation problem monitor the temp of your agar medium. I've noticed that to completly melt my agar I have to bring it up to boiling, but it will stay melted to about 120F. So, I process my agar in the pressure cooker, pull it out and then let it cool with a temp probe stuck under the vessel. When the agar medium cools below 150F I pour my plates and slants. I still will end up with condensate, but not nearly as much.
I have read the input about pouring at a lower temp; my resistance to doing this is I would greatly prefer to sterilize the media in the tubes due to my environment. But I get the feeling I might not have a choice.

fm
 

fratermus

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OK, did a couple more experiments.

Did one where I sterilized the tubes and agar seperately (in the same cooker) and poured the agar after it had cooled substantially. Perhaps a 30% reduction in condensation, thought the droplets are much finer and more evenly dispersed.

Also, I took the tubes that were sterilzed with the agar in them, but uncapped and inverted them in on paper towel in a sanitized container somewhat like Brick mentions for dishes. After 36hrs most of the condensation was gone. flamed the lips and popped on sterile caps. If nothing grows on the media (or if I only lose a couple) then maybe that is a workable solution.
 

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Keep in mind that pyrex and kimax glassware come in "grades". There's a whole spectrum of quality. The most common is "student grade" which is ok but definitely easier to break than the better grades of glassware.

HTH,

M.
 

fratermus

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Did another experiment; poured the liquid media into tubes, stuffed a cotton ball in the ends and pressure cooked. Some slight condensation which evaporated after a couple of days. I "canned' some stoppers in the same batch so when it is time to innoculate and seal I will have sterile stoppers for them.

Will leave them out for a week or so at room temp and see if anything grows on the media; if not maybe this a workable low-condensate process.

Regarding various grades of borosilicate labware, I don't cowboy my glass with rough handling or unwarranted thermal shock so I am not particularly worried. I do wear safety glasses when working with glass, but then again I am a safety geek.
 

Revvy

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I finally got everything together, and plan on starting to make up some slants of bottle cultured pacman this weekend.

The only thing is I can't find glass tubes with stoppers to save my life (flat bottomed or otherwise) I finally bought at the medical supplies store at the medical school where I work some sterile plastic autoclavable tubes...Except they have pointy bottoms...I need to come up with some sort of stand for them...
 

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This is a great thread. :off: Anyone have slants of something other than Wyeast or WhiteLabs yeast they want to swap? I have CL-50, CL-110, CL-600, and Pacman.
 

fratermus

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This is a great thread. :off: Anyone have slants of something other than Wyeast or WhiteLabs yeast they want to swap? I have CL-50, CL-110, CL-600, and Pacman.
Did you harvest CL-110 out of a bottle?

Someday I hope to have enough of a ranch to make swaps a reality.
 
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Schlenkerla

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I look forward to the day when my brewing becomes advanced enough to have a yeast bank.
You can, make slants and use one everytime you brew w/ a new yeast.

Take out about 4 eye droppers from each smack pack before pitching it in your wort. Add this to sterile test tubes and you have the makings for the 1st sample of the bank.

I beefed up my bank by 5-6 strains by going to the LHBS and sorting through the old yeast that was marked down to $2.50 each. The risk is low, the yeast is still sterile and you need to make a starter anyhow.
 
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Schlenkerla

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Today I made new slants in tiny ketch-up bottles (AGAIN!!!). I did this so I could increase my bank of yeasts w/o having to buy more test tubes.



These will be easier to streak and examine yeast growth. Not to mention I can give away yeast if I top them off with sterile wort.

Do you guys usually use agar-agar powder?

I've been wondering why my slants are were kind of soft even when I doubled the amount agar. I made these last night with the powder, and the stuff was still kind of soft. They stayed solid but would spin in the bottle. (Like self-leveling). I looked at the agar package, its got sugar & agar!!! Arrrrrggghhh!!!!

I got the real thing now, Agar-Agar strings. I think they are like seaweed. OMG what a frick'n difference!!!

 
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