Slanting yeast

Homebrew Talk - Beer, Wine, Mead, & Cider Brewing Discussion Forum

Help Support Homebrew Talk - Beer, Wine, Mead, & Cider Brewing Discussion Forum:

This site may earn a commission from merchant affiliate links, including eBay, Amazon, and others.
There is some growth at this moment, I hope I'll be able to select few healthy colonies until Sunday.
 
Well I transfered this yesterday because I wanted the yeast to pitch into a new batch2.
ha2.jpg
 
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.

I am just in the stage of accumulating equipment to start making slants but I have been contemplating adding mineral oil to extend the "shelf life" of the slants. One thing I have been wondering about is sterilizing the tool (dropper, pipette, etc.) that you use to add the mineral oil. Probably over-thinking things but how do you manage this?
 
I am just in the stage of accumulating equipment to start making slants but I have been contemplating adding mineral oil to extend the "shelf life" of the slants. One thing I have been wondering about is sterilizing the tool (dropper, pipette, etc.) that you use to add the mineral oil. Probably over-thinking things but how do you manage this?

I believe the pipette is auto clabeable so you can pressure cook it. Boil it. Star San it during use. Little fire thing to kill more things. Endless possibilities.:)
 
I am just in the stage of accumulating equipment to start making slants but I have been contemplating adding mineral oil to extend the "shelf life" of the slants. One thing I have been wondering about is sterilizing the tool (dropper, pipette, etc.) that you use to add the mineral oil. Probably over-thinking things but how do you manage this?

I don't have an autoclavable pipette/syringe/dropper. I've done it a couple of ways. I was using an oral medicine syringe (good for loading up a several loads of oil at one time - I used around 2-3 ml oil per vial). Cleaned, and then starsanned. I also used a spoon, which you can put in the oven or even flame it, but isn't practical to pour in the opening of the vial. Can make a bit of a mess, but does work. And you have to go back to the primary oil jar multiple times.

I am actually looking into getting something I can boil. Autoclaving take as a long time, with the cooldown. Maybe something like this pipet with pump
 
Thanks for the reply! I like the idea of something "flameable". I might borrow a spoon and bend it into something that is better suited to pouring into the vials. I had that same pipet in my cart at Cynmar but it didn't occur to me to boil it. Maybe a stainless tube that can be used like a pipet. Could flame that (but would the interior get hot enough)? I think I might start without the oil :)
 
very glad that I came across this thread. i have been wanting to culture yeast somehow so I dont have to keep buying fresh yeast for every brew and this seems like the best method. but I do have a question. Say i make 5 slants from one fresh yeast culture. If I want healthy first generation yeast for ever brew will i have to buy fresh yeast after these slants are used up? Or once i make a starter from one of these slants can I then use that starter to inoculate 5 more slants? and essentially never run out of this yeast strain? Or would a slant made from this starter not be considered a first generation yeast?
 
very glad that I came across this thread. i have been wanting to culture yeast somehow so I dont have to keep buying fresh yeast for every brew and this seems like the best method. but I do have a question. Say i make 5 slants from one fresh yeast culture. If I want healthy first generation yeast for ever brew will i have to buy fresh yeast after these slants are used up? Or once i make a starter from one of these slants can I then use that starter to inoculate 5 more slants? and essentially never run out of this yeast strain? Or would a slant made from this starter not be considered a first generation yeast?

Let's say your slant is Batch No. XXX.1, you would make fresh slants from the yeast in your second to last slant from that batch series. Just flame your loop, dip into the yeast in your slant, and rub some of that in a fresh sterile slant, repeat for as many new slants as you would like to create. Then when you're done, use the "mother" slant to make a starter for your next batch. The reason you use the second to last and not the last slant is in case one of those final two is infected. You at least have two to take a peek at and see which is more viable to act as the "mother" slant. The newly created slants are now considered Batch No. XXX.2 and have undergone very little, if any, mutation from the "mother" slant from Batch No. XXX.1.
 
Let's say your slant is Batch No. XXX.1, you would make fresh slants from the yeast in your second to last slant from that batch series. Just flame your loop, dip into the yeast in your slant, and rub some of that in a fresh sterile slant, repeat for as many new slants as you would like to create. Then when you're done, use the "mother" slant to make a starter for your next batch. The reason you use the second to last and not the last slant is in case one of those final two is infected. You at least have two to take a peek at and see which is more viable to act as the "mother" slant. The newly created slants are now considered Batch No. XXX.2 and have undergone very little, if any, mutation from the "mother" slant from Batch No. XXX.1.

awesome! thanks for the info. is there a maximum amount of times you can re-slant like this? Will the yeast eventually get to a point where it is too mutated from the original strain to use?
 
Yes, but there is no definite generation that you can say it will go bad. You can't say "oh that's 1187 Ringwood, only lasts 4 generations before it goes bad."

It is a matter of sanitation, yeast health, and proper storage. If you do all well, you can keep a yeast going, well I won't say indefinetly, but a very long time. I believe some have gone dozens of generations (and are still going). There are threads out there...
 
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 made my agar by this recipe and it set up well (it actually set up in less than 1/2 hour, even before I sterilized it). I inoculated a plate with Bell's yeast 4 days ago but so far there is absolutely no sign of growth. It is stored in the dark at around 75 degrees. I'm thinking that I did something wrong when measuring out the ingredients for the Agar and it is too dry.
 
I sacrificed a plate to test the agar and it is plenty "juicy". Has a wet texture to it. I guess i'll just give it more time.
 
I sacrificed a plate to test the agar and it is plenty "juicy". Has a wet texture to it. I guess i'll just give it more time.

My plates and slants setup within a few hours. If yours is still wet & juicy one of two things likely happened. A) You didn't use enough agar B) the agar didn't fully 'bloom'/disolve with the wort. I had that happen on my first slants, where when dumping the last bit I noticed there was a mass of thick gel on the bottom where most of the agar had ended up.

These days I let the agar bloom in the wort for 15-20min before heating it up initally and never seem to have issues.
 
I made 12 slants yesterday and I just checked them. There is about a 1/4" of gelatinized wort at the bottom. The remainder is all liquid. Any idea what I might have done wrong? Not enough agar agar?

I only ordered 12 vials (more on the way) so I cut the recipe in half. 200ml water, 20g DME and 1.2g agar agar.
 
I made 12 slants yesterday and I just checked them. There is about a 1/4" of gelatinized wort at the bottom. The remainder is all liquid. Any idea what I might have done wrong? Not enough agar agar?

I only ordered 12 vials (more on the way) so I cut the recipe in half. 200ml water, 20g DME and 1.2g agar agar.

Post above you should help you
 
Attached are two pictures of the first starter that I made using a yeast slant. I've made roughly a dozen starters in the past, often right from a White Labs vial. I've heard people talk about pitching starters at high krausen but I've never actually had a lot of krausen on a starter. A few bubbles, maybe some traces of foam but that's it. I use fermcap so I figured that must be it.

NEVER have I seen this much krausen on a starter before. If you're even thinking about making yeast slanting next on your homebrewing adventures, I highly recommend it.

The first picture is yesterday, about 5 hours after I pitched the yeast from the 250ml starter into the 1.1L starter. The second picture is when I got up this morning.

IMG_1199.jpg


IMG_1201.jpg
 
I mash all my starters and have to deal with krausen most of time.. blessed anti-foam :)

Anyone slanted 2 yrs old yeast?
I got 3 WLP vials to isolate @least single colony.. wlp001/004/300. Suppose that wlp300 could be the trouble one.
 
There's now a solid 2" of krausen on the starter. I can't even see the vortex from the stirplate. I'm brewing right now and I started a 90 minute hopstand about 10 minutes ago. In about two hours this thing gets pitched - I may need a blowoff tube. This should be one violent fermentation. I'm glad I'm fermenting this batch in a glass carboy. :)
 
This might have been covered, but how are guys getting slants without hot break? I am using the recipe with 1.5x agar 7x DME 1x Wyeast Nutrient per 100ml water.

Also, my slants have condensation from cooling. There is enough to kind of make drops on top of the media. The media is solid but way juicy. Is this a problem?
 
This might have been covered, but how are guys getting slants without hot break? I am using the recipe with 1.5x agar 7x DME 1x Wyeast Nutrient per 100ml water.

Also, my slants have condensation from cooling. There is enough to kind of make drops on top of the media. The media is solid but way juicy. Is this a problem?

Anybody?
 
You're getting hot break in your agar? I got hot break when I made the wort that I used with the agar to make the slants but none that I can see when I sterilized the slants themselves.

I got that condensation as well. When I inoculated the slants the liquid ran down the sides so that yeast got below the mass of agar. The CO2 generated there pushed the agar plug up (and ultimately out) of the vial. I opened the caps of all of the vials a bit and left them at room temperature for 5 days to dry out. That seems to ahve helped and I will do it from now on as standard practice. Two "samplings" later and I have not experienced that same issue again.
 
This might have been covered, but how are guys getting slants without hot break? I am using the recipe with 1.5x agar 7x DME 1x Wyeast Nutrient per 100ml water.

Also, my slants have condensation from cooling. There is enough to kind of make drops on top of the media. The media is solid but way juicy. Is this a problem?

Lower your quantity of agar. If you're using agar powder, reduce it to 0.5-1.0x and see what your results are. It'll still set up just fine, but allow you a little more time to work with it before it starts setting up. Also, you'll still have hot break no matter what you do, but it does not have any negative impact on yeast viability, stability, and storage time. It's minimal at most compared to the amount seen with an all-grain starter.

You're getting hot break in your agar? I got hot break when I made the wort that I used with the agar to make the slants but none that I can see when I sterilized the slants themselves.

I got that condensation as well. When I inoculated the slants the liquid ran down the sides so that yeast got below the mass of agar. The CO2 generated there pushed the agar plug up (and ultimately out) of the vial. I opened the caps of all of the vials a bit and left them at room temperature for 5 days to dry out. That seems to ahve helped and I will do it from now on as standard practice. Two "samplings" later and I have not experienced that same issue again.

To reduce the chance of "blowouts" make sure to use just enough to cover the entire slant tube from the top edge of the start of the neck-threads to the just below the opposite bottom corner. This will ensure that there is no way CO2 can form entirely below the agar gel and force a "blowout" when you then remove the cap and release the pressure. One way I've found to work great is to buy a cheap 2 tbsp. (30 ml) veterinarian/pediatric liquid medicine syringe or a 2 tbsp (30 ml) turkey injector and use it to accurately draw up the exact same amount I need for each vial and dispense that way before putting them into the pressure cooker. Trial and error with water will tell you how much to use and once you find that amount just mark it with a sharpie. For the popular 6 dram (23 ml) flat bottom vials that a lot of people use, that sweet-spot quantity is about 12 ml.
 
2013_05_20_21_55_36_455.jpg


This look fine to you guys? I got my hands on some Heady so I don't want to blow my chances on culturing this yeast.

The agar set up in a manner than I seem to remember from high school biology so I think I am fine there (using powder) but I might change it up anyway in the name of science.
 
After reading all 47 pages across 2 days, I'm gonna try to liven the Brett talk back up. What I gathered until now regarding Brett slanting is this:

1. Brett prefer pure glucose than other sugars. Anyone tried adding grape sugar (aka dextrose, aka glucose) to their medium recipe? How much?
2. Brett take longer time to propagate. Anyone slanted Brett and can confirm this? Did you let the Brett incubate for a longer time before closing the slants for storage? How long?
3. Brett survive storage time better in room temp then in fridge temp. Can anyone confirm? Did you make wake Brett from fridge successfully?
4. Waking the Brett: Did you step up any differently then Sacch?

If anyone has anything to add, I would be forever in your debt (and my to-be Brett colonies would be as well). This thread has a lot of info about slanting Sacch in general, but very little about Brett. I'd like to change that..
 
Made my first batch of slants on Saturday. They set faster than I was expecting, I think I'll use less agar next time. One thing I'm concerned about though is that there is some sediment in the bottom of nearly every vial. Is this normal or did they pick up some kind of infection before they set?
 
Made my first batch of slants on Saturday. They set faster than I was expecting, I think I'll use less agar next time. One thing I'm concerned about though is that there is some sediment in the bottom of nearly every vial. Is this normal or did they pick up some kind of infection before they set?

It's totally normal

I had sediment in the bottom of all of mine as well. I think it is un-dissolved yeast nutrient.

It's not yeast nutrient. It's actually trub because even LME and DME produce hot break proteins that fall out of suspension. Yeast nutrient is nothing more than basic vitamin salts, i.e. stuff that dissolves rapidly in boiling water.

Keep up the good work guys and enjoy slanting. It's not in your average homebrewer's bag of tricks.
 
It's totally normal

Glad to hear it. I got a bit distracted and let my pressure canner sit for longer than it needed too (like a couple hours longer) when the pressure was coming down before I taped up the slants. Was worried something had creeped in there.
 
Glad to hear it. I got a bit distracted and let my pressure canner sit for longer than it needed too (like a couple hours longer) when the pressure was coming down before I taped up the slants. Was worried something had creeped in there.

There's no way anything can. It's a closed system that was previously inundated with 250F heat and was rendered sterile, not just sanitary.
 
In section #5, when you mention dipping the paper clip into the yeast, you don't mean dip the paper clip back into the yeast each time you submerge it in the slant, right? You just dip it in the yeast once and then into the slant 4 to 5 times. How does it stay sterile exposed to air during the incubation period?
 
In section #5, when you mention dipping the paper clip into the yeast, you don't mean dip the paper clip back into the yeast each time you submerge it in the slant, right?

Nope, that's exactly what I do.

You just dip it in the yeast once and then into the slant 4 to 5 times.

The first time you dip it in, there are very few yeast cells left on the paper clip.

How does it stay sterile exposed to air during the incubation period?

RDWHAHB :mug:
 
Are there any "cheats" for those of us without a pressure cooker? How about a large enamel pot commonly referred to as a "canner". Could I put the vial filled beakers in the canner, filled with appropriate water, and them bake the canner in the oven to get the right temperature? Or would it work on the stove top. Would the plastic lids melt? Thank you.
 
Are there any "cheats" for those of us without a pressure cooker? How about a large enamel pot commonly referred to as a "canner". Could I put the vial filled beakers in the canner, filled with appropriate water, and them bake the canner in the oven to get the right temperature? Or would it work on the stove top. Would the plastic lids melt? Thank you.

You need pressure to achive the heat necessary for sterilization. Otherwise you may get unwelcome critters in your slants.

Dry heat sterilization is possible but takes way longer and at higher temps. Not sure autoclavable plastics could handle it.

You can find decent deals on pressure cookers on Amazon.
 
While at the homebrewing store today I noticed they had a large variety of liquid yeasts that were "past their prime" at discounted prices. Would these yeasts be good for slanting? Would the process outlined in this thread produce a strong yeast, even though it was started with a yeast that was past it's prime?
 
NaymzJaymz said:
While at the homebrewing store today I noticed they had a large variety of liquid yeasts that were "past their prime" at discounted prices. Would these yeasts be good for slanting? Would the process outlined in this thread produce a strong yeast, even though it was started with a yeast that was past it's prime?

Make a starter and you should be fine. I buy this yeast all the time with no problems.
 
While at the homebrewing store today I noticed they had a large variety of liquid yeasts that were "past their prime" at discounted prices. Would these yeasts be good for slanting? Would the process outlined in this thread produce a strong yeast, even though it was started with a yeast that was past it's prime?

If you are going to make slants with them you will be fine, they probably just dont have enough cells for a healthy ferment on their own without a starter. But you will make a starter from your slant so you will be fine. I always go through the past there prime yeast to make slants from and they have all worked fine for me.
 
Back
Top