Slanting yeast

I notice that the author lists bottled water as the water source. Why is this? If the medium is being boiled to such a temp, why is the purified water need? Is the bottled water used for the medium only, or is it also used as the water the fills the pressure cooker too? Thank you.

Hollis, I was wondering if that was the case too for me. The first attempt left me with watery slants, so I upped the agar. Now they seem to hard. I'm wondering if my yeast growth is thin because of that.

Just wanted to share a picture of my 10ml vial that I just innoculated with yeast from my slant..nice krausen forming after 20 hours or so..

tally350z said:

Just wanted to share a picture of my 10ml vial that I just innoculated with yeast from my slant..nice krausen forming after 20 hours or so..

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Would you mind posting your process to get to here? Did you use an inoculation loop to take colonies from a slant and put it in the vial with 10ml of sterile wort?

Also, when filling the vial, do you use canned starter that has been pre-sterilized? Or do you fill the vial, then sterilize it?

HollisBT said:

Would you mind posting your process to get to here? Did you use an inoculation loop to take colonies from a slant and put it in the vial with 10ml of sterile wort?

Also, when filling the vial, do you use canned starter that has been pre-sterilized? Or do you fill the vial, then sterilize it?

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Sure. I usually make about 1000ml of starter wort on the stove top in a sauce pan. Dissolve the DME, than divide it up between about 10 vials 10ml each, and two beakers one with 90 ml of wort and one with 400 ml of wort. Than I autoclave all of the vials and beakers to get sterile wort in each. I usually make 10 vials that way I can have sterile ones handy, and if I let them sit for a few days I can guarantee they are sterile.

When I am ready to make a starter for a beer, such as this one, which is a nut brown ale that I will be brewing thursday. I take a loop from my slant of wlp004, inoculate the 10 ml vial, loosen the cap to let out the co2. Let that go for 24 hours or so, enough time to verify you have an active starter. Pic of the krausen is what you want. Than pitch that 10 ml into the 90 ml for a total of 100ml. Let that go for 24 hours. I'll post pics later. Than you will pitch that into the 400 ml of starter for 500ml total. Than go to 1000ml, and then so on until you have the proper pitching amount.

So do you just select a single colony from the slant to inoculate with? Do you then re-seal the slant to reuse at a later time?


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The first thing I do is take a loop from either the smack pack or White labs vial. Whichever yeast I am using. Than I streak a plate with the loop and let that grow for a few days to maybe a week, than I take a single colony from the plate and streak a slant. Let the slant go for a few days, unsealed. Than I cap it and put it in the fridge. When I want to use the yeast I just pull a full loop from the slant. My slants have some pretty good growth on them. I reseal the vial and put back in the fridge. I haven't gotten to the point were I need to transfer from slant to slant..

Stupid question but do you aerate your tubes or beakers?


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No, I just shake them periodically..

Here is the 100ml beaker that I dumped the 10ml into..about 15 hours in..

I have a yeast strain (WLP022) that I was planning on slanting from a large starter that I made earlier in the week, but now I have a question about it... The starter is completed, and the yeast is rather flocculant and now I am having a tough time getting the yeast to disperse so that I can pour a small sample that I can slant from...

Should I be concerned at all about any selective harvesting that might result from this? Keep shaking/stirring to try to get it back into solution?

Also, I bought an alcohol lamp to use for my slanting sterilization, and it didn't come with any instructions at all... Am I correct in assuming that I just fill it with rubbing alcohol and light the wick?

I know that is a silly question, but my girlfriend will probably get upset if I explode the house.

Have you put the starter in the fridge to cold crash it? If you have than I wouldn't worry about getting the selective yeast from the slurry as most of it would have dropped out. If you haven't than I would do the best you could to get them back in solution. For the alcohol lamp you want to fill the bottom and let the wick soak up the solution. I have heard people who try and pour the solution from the top of the wick and than light it, that's a no no..

tally350z said:

Have you put the starter in the fridge to cold crash it? If you have than I wouldn't worry about getting the selective yeast from the slurry as most of it would have dropped out. If you haven't than I would do the best you could to get them back in solution. For the alcohol lamp you want to fill the bottom and let the wick soak up the solution. I have heard people who try and pour the solution from the top of the wick and than light it, that's a no no..

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I put it in the fridge for a few hours the other day, before I remembered that I hadn't slanted it yet... It's been out of the fridge for about 12 hours. I might put it back on the stir plate to try and get the little suckers swimming again.

Thanks for the input on the lamp, that is pretty much what I figured, but just wanted to be sure that regular isoprop was what I should be using.



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Awesome instructions!
Thank you.

HollisBT said:

Thanks for the input on the lamp, that is pretty much what I figured, but just wanted to be sure that regular isoprop was what I should be using.

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Most instructions will indicate ethyl ethanol (denatured alcohol) for fuel. I ran out and started using isopropyl. It's a lot slower to get the loop red hot, but still works.






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I started using isopropyl but starter running into the problem of the flame turning off, switched to denatured ethanol and now the flame is consistent and much hotter, I'd recommend going this method. Sucks having the fire go off mid slant


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Where do you guys buy the denatured ethanol? I didn't have any problems with the isopropyl the other day, but would still rather use the right stuff.


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Fwiw I am using 91% isopropyl, don't know if that makes a difference or not...


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You can get denatured alcohol at Home Depot. It's shellac thinner and a camp stove fuel. I'll bet you could use Everclear as well.


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Oh, damn. I have a ton of camp fuel lol.


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Not Coleman gas. That's different!


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Oh... That's what I have lol.


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This stuff:


Or the non-toxic option:

If your down on the border. $6 / L

I decided a couple of days ago to brew this weekend and for the first time, I actually had the yeast I wanted slanted and ready to be propagated. I started Wed night with 250ml of sterile wort in a 500ml flask on a stir plate. This morning I stepped up to a little over 1L in a 2L flask. This evening I thought I was going to need a blow off tube for my starter. I have never had such an active starter. I think I got this yeast banking thing working.

Yea, they can be very active..This is nearly 1 hour after pitching my 500ml step into my 1L step..

I am about to start propagating a culture for a high gravity brew, my steps that I'm going to follow are 10ml > 100ml > 1l > 2l.

That should get me the cell count that I need. My question though, is regarding the last step. Should I cool crash, decant, and pitch to 2l? Or would it be ok to pitch the 2 liters of wort on top of the 1 liter of already fermented wort in my 5l flask?


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Best to go ahead and crash and decant.

HollisBT said:

I am about to start propagating a culture for a high gravity brew, my steps that I'm going to follow are 10ml > 100ml > 1l > 2l.

That should get me the cell count that I need. My question though, is regarding the last step. Should I cool crash, decant, and pitch to 2l? Or would it be ok to pitch the 2 liters of wort on top of the 1 liter of already fermented wort in my 5l flask?


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I'll offer my opinion and some thoughts. When stepping up I like to make each step while the culture is in log phase. This allows the yeast stay in reproductive mode and keeps things moving along smoothly. After the final culture is complete allow 8 hours for the yeast to build up its store of carbohydrates then crash (if necessary) decant and pitch. I would also like to offer an alternative step scheme. I have found that starting each step with approximately 35 million cells/ml will achieve the highest yield factor and greater cell doubling (this is also backed up by science). So in the end you have the greatest number of new cells. I have also found that a 6 or 7 fold increase (the European standard) with each step will achieve approximately that 35 million cells/ml/step goal. So consider going from your 10ml initial culture to 70ml > 500ml > 3000 or 3500ml. In my experience the final 3580ml culture will contain approximately 800 billion cells. I do count cells using a hemocytometer and have made this observation several time with a significant margin of error. But according my counts you will be between 700 billion and 900 billion. Please note that very good aeration is necessary to achieve these counts. I set my stir plate so that the vortex reaches all the way to the stir bar and after the initial few minutes the wort looks cloudy due the air that is being dissolved into the media. Some on this forum have suggested that stirring at such speed can damage the yeast but I don't think that is possible. I have certainly never observed any apparently damaged or ruptured cells.

Hmmm, so one vote for crash and one vote to pitch on top haha.

I'd prefer just to pitch it without the crash, for a few reasons... Mainly for time constraints, but also as previously noted to keep the yeasts at their most active.

My logic on the dilution (I am using canned starter wort) is that it would still have the same amount of fermentables in it, just at a lower concentration. Does that seem logical? According to the calculator I'm using the lower gravity that I would be starting at should still yield the cell count that I am aiming for.


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Hollis how did it turn out for you?

Pitched my 1.5L starter in the wort and it hit FG within 4 days and tastes great.

tally350z said:

Hollis how did it turn out for you?

Pitched my 1.5L starter in the wort and it hit FG within 4 days and tastes great.

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Well I ended up not brewing the beer (yet). But I decided not to crash, and added the last addition on top of the 1l starter as it was starting to slow down.

The last step seemed to take a little while to complete (part of the reason for delaying the brew), which in retrospect is probably due to the lower concentration of sugars in the starter wort.

It did eventually finish though, and is currently in the fridge to let it clarify, I'll probably brew the beer next week.

This was my first propagation from a slant, and I am more than happy with it! Used about a third of one slanted tube and ended up with what looks to be a very healthy sized pitch of yeast. Thanks to everyone here who has helped with my excessive amount of questions lol.


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My most recent slanted yeasts have developed some condensation on the inside if the tubes, with the culture already in them...

Will these be ok? I am assuming this is weather related?




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HollisBT said:

My most recent slanted yeasts have developed some condensation on the inside if the tubes, with the culture already in them...

Will these be ok? I am assuming this is weather related?

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Are they sealed shut already? Or open and being allowed to breathe (i.e. newly inoculated)? If they are shut, then this is totally normal and it's the agar agar media perspiring and releasing water. You may want to move them to a cooler part of your fridge, to help reduce that water loss. I keep my culture tubes right by the fridge vent on the top shelf of my fridge where the cool air enters the lower chamber. Another great place may be the crisper drawer at the bottom of the fridge (heat rises after all). Once the agar agar media dries out completely, the viability of the culture begins to reduce dramatically.

They were inoculated about 10 days ago, I noticed it when I went to deal them and move them to the fridge. So they have been capped loosely for the past several days. Should I seal them and move to fridge?


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HollisBT said:

They were inoculated about 10 days ago, I noticed it when I went to deal them and move them to the fridge. So they have been capped loosely for the past several days. Should I seal them and move to fridge?

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If that is a fairly active yeast and from a very active fermentation, then a little respiration is totally normal. Fermentation is an exothermic reaction that releases heat and therefore cause water vapor along with CO2 to be released. Look again at some of your higher gravity batches of beer and look closely at the tube coming into the bottom of your airlock, they commonly have condensation built up from all that water vapor.

Seal them and move them to the fridge. I never allowed my slants to be inoculated in a warm weather environment to begin with. I always slanted and immediately placed them into the fridge. Never had a problem, I just had to uncap them less frequently to release CO2 as the fermentation in the tubes was not extremely active.

I saved a couple of yeasts on slants over the past couple months, but had yet to actually use one. Well I finally used my slanted london ale yeast wlp013 a recently, and I have to say it went better than I expected!

I took a loop of yeast from the slant and stuck it in 10ml of wort, then built it up to 70ml, 500ml, and then 3 liters, each after 24 hours, just dumping one into the next. My brew day got delayed so I ended up cold crashing it for a few days.

Well I finally pitched the yeast, there was 1 lag day, then 4 days later its done. It was one of my more vigorous fermentations, moreso than the first time I used wlp013. It also had higher attenuation, and went from 1.072 to 1.010. Very impressed, should make for a great IPA. Looks like slanting is the way to go!

thefost said:

It was one of my more vigorous fermentations, moreso than the first time I used wlp013. It also had higher attenuation, and went from 1.072 to 1.010. Very impressed, should make for a great IPA. Looks like slanting is the way to go!

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I've seen the same thing in my 1 and only fermentation from a slant. This was the only time ever that I had a need for a blowoff tube with a Speidel 30L fermenter. It was a 5 gallon 1.060 batch with my normal temperature controlled fermentation. I've got a 1.083 in there right now that came nowhere close to needing a blowoff. As I continue to build up my library, I am looking forward to more opportunities to use my slants. Slanting saves me a few $$, but I am really using it in order to keep a large variety of yeasts on hand.

Took my first stab at this. I think I need to redo the slants. They don't seem to be setting. The bottom of the vials had about a half inch of solid, and the rest is like loose gelatin.
I think I need to use more agar powder in the mix than 2.5 grams