Slanting yeast
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jason.mundy
BIAB Purist
I had a tube of WLP830 I wanted to slant, but didn't have any slants ready yet. My solution was to not pitch the entire tub into my starter. I saved back about an 1/8th of the vial. I assume it will be good as long as the expiration date on vial.
I thought this would be better than grabing a sample from a starter because the vial should be bacteria free when it was filled at White labs.
Good idea. In that same line of thinking I could probably put a small amount of Wyeast in a vial and do the same. Thanks!
supermoth
Well-Known Member
Awesome tutorial, and thanks for the idea. Here are my slants.
For those of you who don't have all the equipment ready to do this, I highly suggest you bribe someone who has access to a microbiology lab to do it for you. I did this at my workplace (on lunch, of course!
) and it was a snap.
For those of you who don't have all the equipment ready to do this, I highly suggest you bribe someone who has access to a microbiology lab to do it for you. I did this at my workplace (on lunch, of course!
) and it was a snap.
Will plastic viles work for this? I've searched this thread but cannot seem to find anything mentioning the use of plastic viles. I have around 40 or so White Labs viles from previously used yeast. I would like to use them but am not sure if the plastic can hold up to the heat from the pressure cooker. They seem like pretty heavy duty plastic to me. The viles are also a little bit bigger. Has anyone here slanted by using plastic viles?
-TPM-
Well-Known Member
The White Labs viles aren't any good, they melt a bit and turn white so you can't see through them anymore.
-TPM-
Well-Known Member
NP. If you want a cheap source let me know and I'll pm you the site where I got mine (~50c each)
DrawTap88
Well-Known Member
TPM, can I please get the site too? I need to get a few more.
Awesome tutorial, and thanks for the idea. Here are my slants.
For those of you who don't have all the equipment ready to do this, I highly suggest you bribe someone who has access to a microbiology lab to do it for you. I did this at my workplace (on lunch, of course!
Aren't you worried about contamination from your lab? This sounds like a cool project to do at home, but I don't think I'd run the risk of picking up something from the lab or hazardous chemicals. I suppose it depends on what your lab works on. Nice plating, by the way.
Regarding this protocol, wouldn't it be better to use a high proof alcohol, e.g. Everclear, instead of sanitizer? That's what I've done in the lab. Dip the loop, aka paper clip, into the Everclear, flame, and then wave back and forth to cool.
OP
OP
Sure, you could, but the idea of having a bunch of alcohol right next to an open flame gave me the willies. SWMBO would diapprove of me burning the house down while culturing yeast.
Good point, but I bet that if I ever attempt this, I'll be too lazy to dilute and will just use straight Everclear.
Another thought, if you had access to sterile tubes/plates, could you just pressure cook a bottle containing all of your agar agar to sterilize and then pour plates or pour into tubes? I'm guessing that tubes would be better for long term storage than plates in the fridge; you wouldn't want your fridge smelling like yeast. I suppose that you could sterilize all of your empty tubes first, sterilize your agar agar separately, and then transfer. Food for thought.
-TPM-
Well-Known Member
That's actually how they recommend doing plates in 'Yeast - The Practical Guide to Beer Fermentation'. You could do tubes that way too, but it would be harder (pouring hot agar into a tube aseptically...). Yes, tubes are for long term, plates are to get pure cultures and short term.
jason.mundy
BIAB Purist
What is the concern about condensation? Isn't it sterile as well?
I could guess that it would allow your yeast colonies to mix. Is that it?
I could guess that it would allow your yeast colonies to mix. Is that it?
wardenwheat
Well-Known Member
I may have missed this, but do you store your un-innoculated slants in the fridge too, or at room temp somewhere?
jason.mundy
BIAB Purist
I keep mine out at room temp. That way I can see if anything starts to grow on them before I use them. Make sure the lids are screwed on tight to avoid the slants drying out.
In the lab, agar plates/tubes are kept in the fridge to minimize any chance of contamination. While they should be mostly sterile and therefore fine at room temp, I'd still keep them in the fridge. Be sure that they're sealed tightly because they can dry up in the fridge too.
OP
OP
It's time to re-slant my library (some of my slants are well over a year old at this point, and I have some two year old expired yeast vials I got cheap that I need to slant as well). I went to Home Depot yesterday and picked up some boards to build vial racks to incubate the old slants with sterile wort. I plan to let them incubate while I'm gone at NHC, and I'll re-slant all of my yeast strains when I get back. I'll post pics of the racks once I get them built.
mn_brew
Well-Known Member
I tried slanting yeast for the first time on some Wyeast 3787 Trappist High Gravity. It was extremely easy and now I have five slants of an awesome yeast. If you've been thinking of trying this, do it.
wardenwheat
Well-Known Member
My slants looked great after setting up, but have started to turn watery after a couple weeks. I used the proportions mentioned on the first page with pharmaceutical grade agar. Any suggestions??
OP
OP
Possible the mix settled before you poured it? I had that happen with one batch half went to water and had to be tossed. If they are all watery you need more agar.
RJSkypala
Well-Known Member
How do these vials look for slanting?
http://cgi.ebay.com/72-pcs-Clear-4-...aultDomain_0&hash=item1e566a071b#ht_500wt_922
http://cgi.ebay.com/72-pcs-Clear-4-...aultDomain_0&hash=item1e566a071b#ht_500wt_922
RJSkypala
Well-Known Member
I contacted the seller to ask about that.
If they are autoclaveable, are they a good size?
If they are autoclaveable, are they a good size?
OP
OP
Size sounds OK assuming they are autoclaveable.
bv805
Member
Whats the optimal temperature for storing the slants after incubation..I just got a mini fridge for my yeast and its sitting at about 33 F
OP
OP
bv805
Member
33 is right where I try to keep mine. As close to freezing without freezing.
Ya thats what I thought, thanks for the reply and keeping this thread going you've taught me alot.
Brings back memories of my microbiology lab last year, except we were playing with bacteria instead of yeast...wonder if my microbio professor would like to sponsor a "research" project on culturing yeast from bottles
pngaudioguy
Member
First, I want to say I finally got around to registering instead of just lurking because of this thread. Thanks!
I made my slants Saturday evening, and only about half of them set. I didn't think to do the math for myself. Somewhere along about page 8, I found this.
Sacc - you may want to modify your first page to include your recipe so that those of us who go and get excited and start making things before reading the whole thread don't get burnt Awesome info on this site. Thanks!
Jason - those are very sharp looking vials. I will be copying the food coloring on round two this afternoon, with more agar...
Also, just to be safe, I think I'll make sure the slant mix gets heated a little more and stir with the pipet before each sample collection. 5mL calibrated pipettes make simple consistent fills. I also grabbed a test tube rack that is autoclaveable to make sure nothing falls over, etc. I'll grab a picture and post the cynmar number this afternoon.
I made my slants Saturday evening, and only about half of them set. I didn't think to do the math for myself. Somewhere along about page 8, I found this.
Sacc - you may want to modify your first page to include your recipe so that those of us who go and get excited and start making things before reading the whole thread don't get burnt
Awesome info on this site. Thanks!Jason - those are very sharp looking vials. I will be copying the food coloring on round two this afternoon, with more agar...
Also, just to be safe, I think I'll make sure the slant mix gets heated a little more and stir with the pipet before each sample collection. 5mL calibrated pipettes make simple consistent fills. I also grabbed a test tube rack that is autoclaveable to make sure nothing falls over, etc. I'll grab a picture and post the cynmar number this afternoon.
pngaudioguy
Member
As promised, here's a picture of the rack I used, available here:
http://www.cynmar.com/Staging/item_detail.aspx?ItemCode=15025994
It says it's only autoclaveable to 250*F, so I kept the pressure between 12 and 13 to make sure it didn't melt. (15 PSI is required to hit 250, so I figure that's a pretty safe margin.)
http://www.cynmar.com/Staging/item_detail.aspx?ItemCode=15025994
It says it's only autoclaveable to 250*F, so I kept the pressure between 12 and 13 to make sure it didn't melt. (15 PSI is required to hit 250, so I figure that's a pretty safe margin.)
bv805
Member
As promised, here's a picture of the rack I used, available here:
http://www.cynmar.com/Staging/item_detail.aspx?ItemCode=15025994
It says it's only autoclaveable to 250*F, so I kept the pressure between 12 and 13 to make sure it didn't melt. (15 PSI is required to hit 250, so I figure that's a pretty safe margin.)
looks good just ordered those racks and tubes. i previously had morebeers tubes but they were way to small
jason.mundy
BIAB Purist
@pngaudioguy - That's a nice looking rack that holding the vials. I'm not sure it would fit in my 16qt pressure cooker.
Did you need to put anything under it to keep it from touching the bottom of the cooker?
I usually crumple up some aluminum foil to go under my pyrex measuring cup to act as a trivet.
Did you need to put anything under it to keep it from touching the bottom of the cooker?
I usually crumple up some aluminum foil to go under my pyrex measuring cup to act as a trivet.
SpaceJunk
Active Member
Another container that works great for your cultures is Ball or Mason brand jelly jars. They have a widemouth opening like regular jars but are more squat and only a couple inches tall. They stack nicely and the jars give you a lot of surface area on the bottom for your agar yeast culture. When you wish to make a starter from them you can use an inoculation loop to scrape some yeast from the culture and swish it into your starter flask.
pngaudioguy
Member
Jason - I set the rack directly on top of the provided canning insert. I figure the goal is to keep the glass off the metal, which the rack does just fine.
I have a 23 qt, but it appeared to be the same diameter as the 16 qt. You could cut out a piece of cardboard at 4.25" x 9.75" (outside dimensions) and see if it would fit.
I have a 23 qt, but it appeared to be the same diameter as the 16 qt. You could cut out a piece of cardboard at 4.25" x 9.75" (outside dimensions) and see if it would fit.
kappclark
Well-Known Member
Can you get the slant's pre-filled instead of 'rolling your own' ??
