Isolated Yeast (Tree House): How to Identify and Characterize?

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HopsAreGood

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sorry, I don’t understand. What does this tell you?
Agreed...I’ve been following this thread for quite a while and have a very good understanding of all the pertinent details so far. I get a bit lost though with all the scientific babble..I mean that in the nicest possible way.
 

TheHairyHop

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Gotcha, I didn’t realize you were thinking along that line (dry yeast contamination). I don’t think that’s very likely, at least at the scale we are going to pick up on.

My thinking was to see how “clean” this process would be using a commercial beer with a known yeast. Like, I’d you plated the dregs and PCR’d 10-20 colonies, would they all be identical?
Hmm, yea. Both might be necessary in order to rule out any outside introduction. In the case you were talking about, yea, Bells might be a great example.

Apologies for the skepticism, being doubtful of a result is definitely the first reaction I have, probably because of the conservative nature of my field. I guess that I'm just stuck on the WB-06 being diastaticus. Maybe I'm ignorant, but how reliable would a killer yeast be at suppressing the growth of an STA-1 positive yeast? From my understanding and just familiarity with biology, I can't imagine it's a 100% success rate, yet after countless batches, I haven't heard of TH beers exploding, gushing, or even over carbonation. If a diastaticus yeast was being used, I would assume they'd centrifuge it out, but it's being found in bottles 🤷‍♂️
 

Eltes

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Hmm, yea. Both might be necessary in order to rule out any outside introduction. In the case you were talking about, yea, Bells might be a great example.

Apologies for the skepticism, being doubtful of a result is definitely the first reaction I have, probably because of the conservative nature of my field. I guess that I'm just stuck on the WB-06 being diastaticus. Maybe I'm ignorant, but how reliable would a killer yeast be at suppressing the growth of an STA-1 positive yeast? From my understanding and just familiarity with biology, I can't imagine it's a 100% success rate, yet after countless batches, I haven't heard of TH beers exploding, gushing, or even over carbonation. If a diastaticus yeast was being used, I would assume they'd centrifuge it out, but it's being found in bottles 🤷‍♂️
I listened to this webinar (https://www.edudip.com/en/webinar-recording/20e688bb-239f-4041-9683-4a0931acd761) from Fermtis the other week and at the end there was an off-topic question about diastaticus yeasts where the Fermentis guy giving the webinar remarked that diastaticus doesn't equal hyper attenuator. He explicitly gave WB06 as an example of a diastaticus yeast which is not hyper attenuating.
 

TheHairyHop

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I listened to this webinar (https://www.edudip.com/en/webinar-recording/20e688bb-239f-4041-9683-4a0931acd761) from Fermtis the other week and at the end there was an off-topic question about diastaticus yeasts where the Fermentis guy giving the webinar remarked that diastaticus doesn't equal hyper attenuator. He explicitly gave WB06 as an example of a diastaticus yeast which is not hyper attenuating.
Very interesting. Thanks for the link!
EDIT: I can't get it to load, but it might just be Chrome
 

Eltes

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I listened to this webinar (https://www.edudip.com/en/webinar-recording/20e688bb-239f-4041-9683-4a0931acd761) from Fermtis the other week and at the end there was an off-topic question about diastaticus yeasts where the Fermentis guy giving the webinar remarked that diastaticus doesn't equal hyper attenuator. He explicitly gave WB06 as an example of a diastaticus yeast which is not hyper attenuating.
Very interesting. Thanks for the link!
EDIT: I can't get it to load, but it might just be Chrome
Actually it seems they took down the recording again. Luckily I recorded it myself too. I uploaded the video here:
The password to watch it is "HomeBrewTalk".
 
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isomerization

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I listened to this webinar (https://www.edudip.com/en/webinar-recording/20e688bb-239f-4041-9683-4a0931acd761) from Fermtis the other week and at the end there was an off-topic question about diastaticus yeasts where the Fermentis guy giving the webinar remarked that diastaticus doesn't equal hyper attenuator. He explicitly gave WB06 as an example of a diastaticus yeast which is not hyper attenuating.
See this link: The mysteries of diastatic brewing yeast | Suregork Loves Beer

I didn’t see WB-06 mentioned, but I wonder just how actively expressed the STA-1 gene is for WB-06.
 

suregork

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GRBC

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sorry, I don’t understand. What does this tell you?
My best guess is it means you took a slurry of s04, t58 & wb06, streaked then onto Plates, took two colonies of each and compared the DNA markers. S04 & T58 appears to be the same and the wb06 is unreadable. Is that right? If so, I guess it’s. “Duh!” from me. Of course two colonies of yeast from the same packet are identical! What did you expect?
 

Northern_Brewer

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My best guess is it means you took a slurry of s04, t58 & wb06, streaked then onto Plates, took two colonies of each and compared the DNA markers. S04 & T58 appears to be the same and the wb06 is unreadable. Is that right?
Pretty much, if you mean slurries of each strain grown separately, grown on separate plates.

If so, I guess it’s. “Duh!” from me. Of course two colonies of yeast from the same packet are identical! What did you expect?
It's not duh at all. It's proving two things haven't happened :

1) Contamination of the PCR - which does happen, although it's less of the problem for colony PCR like this where there's many copies of the target DNA. It's a real problem if you are eg looking for a few copies of virus genetic material in a messy sample of eg human blood or sputum. Control samples are always good.

2) It's not at all obvious that "two colonies of yeast from the same packet are identical" - in fact there's quite a bit of evidence that in the past dry yeast have not been particularly pure - back in 2014 Chris Giles of Surebrew was reporting finding up to 5 strains in US-05 (probably just flocculation mutants) and there were dark rumours about the purity of US-05 among some commercial brewers in 2017-18. Chris also suggested that Notty was 70% lager yeast and Ed Wray found strains in Notty that grew differently on WLN medium but both grew at 37C (traditional test of ale vs lager yeast).

So no, it's not obvious that one would necessarily find that the banding is the same because "two colonies of yeast from the same packet are identical". Although my personal suspicion is that after the lawsuits over diastatic contamination, all yeast companies probably cleaned up their production strains, so it may not be possible to reproduce the above results with modern US-05 and Notty. Still worth looking at a bunch of colonies though - even better if one could find some "old" Notty in a cupboard somewhere.

Me, I've just got a bit of a crush on how pretty those ladders are, having used old-school plasmid digests for ladders back in the day...
 

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You’re right. It is better to test and know than to assume. I guess I was just hoping that it was a more exciting announcement - that the yeast blend in Julius is confirmed. I’m sure that’s coming eventually.
 

TheHairyHop

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2) It's not at all obvious that "two colonies of yeast from the same packet are identical" - in fact there's quite a bit of evidence that in the past dry yeast have not been particularly pure - back in 2014 Chris Giles of Surebrew was reporting finding up to 5 strains in US-05 (probably just flocculation mutants) and there were dark rumours about the purity of US-05 among some commercial brewers in 2017-18. Chris also suggested that Notty was 70% lager yeast and Ed Wray found strains in Notty that grew differently on WLN medium but both grew at 37C (traditional test of ale vs lager yeast).
Yea, this is kind of what I've been thinking. I just think of all the hubbub around White Labs and their issues, and I don't entirely believe that the yeast coming out of TH's conical is pure. Not that I don't believe it could be a blend, I would just want to know what a 500g brick of S-04 looks like after a generation or two
 

GRBC

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Yea, this is kind of what I've been thinking. I just think of all the hubbub around White Labs and their issues, and I don't entirely believe that the yeast coming out of TH's conical is pure. Not that I don't believe it could be a blend, I would just want to know what a 500g brick of S-04 looks like after a generation or two
The brewery is not sterile, so I would guess that there are some non-zero, but trivial number of uninvited yeast/bacteria in the conical. But even if there is cross-contaminated yeast in a brick of S-04, I would guess the amount is so infinitesimally small that the odds of culturing and selecting that colony from yeast cultured from a can of Julius are extremely low. After all the brewer goes to great lengths In terms of pitch rate, pH, temp, etc. to favor of his preferred fermenter.

tl;dr - IMO if you get three unique strains from a sterile streaking of a can of Julius, it is more likely that they used three strains than one or two rando’s showed up in the can.
 
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mcoman

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Over the past 2 weeks or so I have read through the thread and its full on great information. I have tried my hand at making a NEIPA several times with results from disappointing to good, but never anything really amazing. I have some ideas to try on my next 5 gallon brew this weekend. Let me know what you guys think.

For a grain bill I'm going to try:
Briess 2 row
20% carafoam
7% Vienna
5% C20
3% GNO

I have been doing a 60-75 min mash but I am going to shorten that up to 45 min at 156 to increase the maltiness

For hops a mix of El dorado and moasic probably in a 2:1 ratio. I'm not set on the amounts but probably something like 1oz in the mash, no boil hops 3-4oz in the whirlpool around 140.

Cool to about 78 (my ground water is to dam warm this time of year to get much below that) and under pitch some 1318 without aerating. Leave the beer warm for about 12 hours until fermentation starts than cool to 64 and hold it there until just before its finished, soft crash to 60, closed transfer into a keg with 9oz of dry hop per Scott Janish filter tube method.

Dry hop for 2 days at 60, closed transfer to a serving keg with 1L of kräusen made with a can of propper starter canned wort and some WLP400 that I collected from a belgian wit I made earlier in the spring. Let everything come up to 70 for a week than cold crash and serve.

I am hoping I can get some of the belgian flavors from the WLP400 kräusen, while it scrubs any oxygen from the transfer after dry hopping.
 
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RTE

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I don't mean to be a Debbie downer but I'd recommend locking down 1 basic NEIPA recipe. Work with 1-2 grains and 1-2 types of hops. Dial in your water. Perfect your dry hopping process, closed transfers, and fermentation temperatures. Once you make a non-disappointing beer 2 times or so in a row start experimenting with multiple yeast strains or other variables.

So for example in your above recipe, those 4 types of specialtly grains are only going obfuscate what's going on with the yeast and hops. Ideally you'd work from some simple recipe that you've mastered and the only change you make is adding the belgian yeast. That way it will be really easy to tell if you added to much belgian yeast or too little and if you successfully coaxed out any "biotransformation".
 

brewpharm Hill

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@robopp how have your trials been? I remember you said you have come really close with the combo. I'm going to take another stab at the yeast combo next weekend. Going to use Mr. Malty for my calcs and I have a better scale to use. No aerating or rehydration and no yeast nutrient.
 

HopsAreGood

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Any updates? Curious how this turns out.
I’ll be kegging it tomorrow 13 days from brew day. I dry hopped it with 8oz of mosaic so hopefully I’ll be able to notice whatever the 71B is bringing to the table. Fermentation was pretty typical, nothing abnormal but it did have a very pleasant smell that was a bit different than usual. Very tropical, definitely fruity and estery. I’m going to quick carb this one so I should be able to have a taste in a day or two. I’ll keep you updated.
 

HopsAreGood

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Ok, so I just transferred it and poured a glass to taste. It’s completely flat with no carbonation but it definitely tastes very different than any beer I’ve ever brewed. It’s still beer, but there’s absolutely a fruity wine kind of thing going on. It’s kind of hard to explain and I’m excited to see what happens as it carbs up and conditions over the next week or so. Some of the sips I take taste very much like a fruity white wine, and then other sips just taste like this sort of fruit salad medley that does in fact remind me a little bit of treehouse. I can say for sure that this is not the magic that treehouse is doing, but it definitely does come pretty close in terms of aroma, mouth feel, and overall character. If nothing else it’s definitely a very interesting experiment and I think this beer is going to turn out really really good. It started at 1.072 and ended at 1.014. I’ll give an update after it’s carbed and a little bit more conditioned.

One thing I’ll add is that while I don’t expect to get extremely defined hop character 10 minutes after kegging the beer, there is no sign of the 8 ounce mosaic DryHop. It’s straight yeast Character which is primarily what I get from the core treehouse beers. I have a hard time finding any hops in them, to me they just have that fruity bubble gummy Melon like yeast thing going on...So definitely similar in that regard.
 
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TBryerton

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View attachment 685096View attachment 685097Ok, so I just transferred it and poured a glass to taste. It’s completely flat with no carbonation but it definitely tastes very different than any beer I’ve ever brewed. It’s still beer, but there’s absolutely a fruity wine kind of thing going on. It’s kind of hard to explain and I’m excited to see what happens as it carbs up and conditions over the next week or so. Some of the sips I take taste very much like a fruity white wine, and then other sips just taste like this sort of fruit salad medley that does in fact remind me a little bit of treehouse. I can say for sure that this is not the magic that treehouse is doing, but it definitely does come pretty close in terms of aroma, mouth feel, and overall character. If nothing else it’s definitely a very interesting experiment and I think this beer is going to turn out really really good. It started at 1.072 and ended at 1.014. I’ll give an update after it’s carbed and a little bit more conditioned.

One thing I’ll add is that while I don’t expect to get extremely defined hop character 10 minutes after kegging the beer, there is no sign of the 8 ounce mosaic DryHop. It’s straight yeast Character which is primarily what I get from the core treehouse beers. I have a hard time finding any hops in them, to me they just have that fruity bubble gummy Melon like yeast thing going on...So definitely similar in that regard.
Thanks for the update - keep us posted.

I keep going back to fact that Nate is an advocate of process. If it’s true that he adds hops prior to fermentation ending for beer that uses the ‘house yeast’ and he does not for others (such as Bright) there has to be a good reason. And I’m convinced it has something to do with blending yeast.
 

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"Our house yeast provides juicy fruit gum and melon overtones throughout."

 

SanPancho

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WB-06 has the deletion in the STA1 promoter, so STA1 is not expressed very actively. It can still ferment dextrins, but it happens very slowly.
Or not. brew with 95/3/2 blend finished at 1.014. same week? Coronavirus shut down. Got left in the tank. After a Month at ambient it’s at 1.008. Two months later? Dunno. Stopped trying to sample as now tank is at 20psi and sample comes rocketing out as pure foam.

doesnt seem like its the s04 or the t58.......
 

beervoid

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View attachment 685096View attachment 685097Ok, so I just transferred it and poured a glass to taste. It’s completely flat with no carbonation but it definitely tastes very different than any beer I’ve ever brewed. It’s still beer, but there’s absolutely a fruity wine kind of thing going on. It’s kind of hard to explain and I’m excited to see what happens as it carbs up and conditions over the next week or so. Some of the sips I take taste very much like a fruity white wine, and then other sips just taste like this sort of fruit salad medley that does in fact remind me a little bit of treehouse. I can say for sure that this is not the magic that treehouse is doing, but it definitely does come pretty close in terms of aroma, mouth feel, and overall character. If nothing else it’s definitely a very interesting experiment and I think this beer is going to turn out really really good. It started at 1.072 and ended at 1.014. I’ll give an update after it’s carbed and a little bit more conditioned.

One thing I’ll add is that while I don’t expect to get extremely defined hop character 10 minutes after kegging the beer, there is no sign of the 8 ounce mosaic DryHop. It’s straight yeast Character which is primarily what I get from the core treehouse beers. I have a hard time finding any hops in them, to me they just have that fruity bubble gummy Melon like yeast thing going on...So definitely similar in that regard.
That overly fruity thing is what I got too, it will fade away over time and develop into something that I wasn't a big fan of. I used much less then you though.
Thanks for the update, curious to read how it develops.
 

HopsAreGood

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That overly fruity thing is what I got too, it will fade away over time and develop into something that I wasn't a big fan of. I used much less then you though.
Thanks for the update, curious to read how it develops.
How much did you use? What was the yeast breakdown?
 

HopsAreGood

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I dont know anymore, more like 10%. Do you still have an full 71b bag lying around? I could weigh out my opened one that I still got
No, it doesn’t really matter. I was just curious if you remembered. I did 7g s-04 and 3g 71b. If I do this again I may do the whole 11.5g of s-04 and the whole 5g of 71b. That way it would still be 70% and 30% but it would be more of a substantial pitch so perhaps the ester production would be less pronounced. Although I really have no idea what this is going to actually turn into so I’ll have to wait and see.
 

beervoid

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No, it doesn’t really matter. I was just curious if you remembered. I did 7g s-04 and 3g 71b. If I do this again I may do the whole 11.5g of s-04 and the whole 5g of 71b. That way it would still be 70% and 30% but it would be more of a substantial pitch so perhaps the ester production would be less pronounced.
Just weighed the empty bag I would say I pitched about 1gram maybe 1.5.
 

TheHairyHop

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Or not. brew with 95/3/2 blend finished at 1.014. same week? Coronavirus shut down. Got left in the tank. After a Month at ambient it’s at 1.008. Two months later? Dunno. Stopped trying to sample as now tank is at 20psi and sample comes rocketing out as pure foam.

doesnt seem like its the s04 or the t58.......
Yea, slowly but surely is still something. TH beer gets traded around too much to not be hearing about exploding cans. Could the killer yeast have a 100% success rate? It doesn't actually kill the yeast right? So, I would imagine that the occasional gusher is inevitable. I don't know. Maybe I'm thinking of this wrong...
 

Clyde McCoy

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Summary of PCR results so far -

Screen Shot 2020-06-16 at 2.35.04 PM.png

Screen Shot 2020-06-16 at 2.35.11 PM.png

Screen Shot 2020-06-16 at 2.35.21 PM.png


If you have access to TH beers and are willing to ship (ideally older cans), PM me and I'll test them side-by-side.
 

SanPancho

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Yea, slowly but surely is still something. TH beer gets traded around too much to not be hearing about exploding cans. Could the killer yeast have a 100% success rate? It doesn't actually kill the yeast right? So, I would imagine that the occasional gusher is inevitable. I don't know. Maybe I'm thinking of this wrong...
so, the big argument here is that "killer" is a misnomer, and it doesnt always work on every strain. fair point. but im thinking that it will only have to inhibit wb06 enough, not "kill", to stop it from getting active and going into starvation mode-- which is where you probably start to see sta1 genes getting turned on to start munching dextrin.

not to mention i find it hard to think they wouldnt have figured this out over the years if it DIDNT work and you did get gushers. its been known for a long time now that wb06 is diastaticus. (although i seemed to miss that somehow....)
 

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So I carbed it up and this beer is SOFT and THICK on the palette. It still has that very fruity wine like thing going on but it’s very enjoyable to drink. My best guess is that the 71B is really contributing to the mouthfeel due to the high levels of glycerol it produces. I think I’ll try playing with some different combos going forward...both yeast combos and ratios. My next brew is going to be with omegas version of 1318 so maybe I’ll try that whole pack and 2 or 3 grams of 71B.
 
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isomerization

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View attachment 685299So I carbed it up and this beer is SOFT and THICK on the palette. It still has that very fruity wine like thing going on but it’s very enjoyable to drink. My best guess is that the 71B is really contributing to the mouthfeel due to the high levels of glycerol it produces. I think I’ll try playing with some different combos going forward...both yeast combos and ratios. My next brew is going to be with omegas version of 1318 so maybe I’ll try that whole pack and 2 or 3 grams of 71B.
Okay you’ve inspired me to do a split batch with 1318 and 71B together and just 1318 alone!
 

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View attachment 685299So I carbed it up and this beer is SOFT and THICK on the palette. It still has that very fruity wine like thing going on but it’s very enjoyable to drink. My best guess is that the 71B is really contributing to the mouthfeel due to the high levels of glycerol it produces. I think I’ll try playing with some different combos going forward...both yeast combos and ratios. My next brew is going to be with omegas version of 1318 so maybe I’ll try that whole pack and 2 or 3 grams of 71B.
Beer looks beautiful! Which is omegas version of 1318? Haven't even used 1318 myself yet though lol. Also - when you pitched 70% S04 and 30% 71B, did you rehydrate at all or just measure out the grams and sprinkle on top? Do you aerate wort prior to pitching as well? Just trying to see how you pitched this. 71B definitely has me intrigued though especially if this adds the mouthfeel you are stating here. Keep us posted on how this conditions as well as whether your mosaic starts to shine or if it remains a yeast ester machine!
 

HopsAreGood

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Beer looks beautiful! Which is omegas version of 1318? Haven't even used 1318 myself yet though lol. Also - when you pitched 70% S04 and 30% 71B, did you rehydrate at all or just measure out the grams and sprinkle on top? Do you aerate wort prior to pitching as well? Just trying to see how you pitched this. 71B definitely has me intrigued though especially if this adds the mouthfeel you are stating here. Keep us posted on how this conditions as well as whether your mosaic starts to shine or if it remains a yeast ester machine!
Omega’s version is British ale V or OYL-11...same thing as 1318 or imperial juice. My local Homebrew store carries omega and I’ve always been happy with all of their yeasts including British ale V. For this batch I weighed out 7 grams of s-04 and 3 grams of 71B and I did not rehydrate. I have a small anvil pump and the simple act of pumping it into the fermenter gets it nice and frothy, thats all I really do to oxygenate. I fill 1/3 of my fermenter with wort then pitch in all of the dry yeast, then fill the remaining 2/3 right on top of it to mix it in. 12 hours post pitch and it was chugging away.
 
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Noob_Brewer

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Omega’s version is British ale V or OYL-11...same thing as 1318 or imperial juice. My local Homebrew store carries omega and I’ve always been happy with all of their yeasts including British ale V. For this batch I weighed out 7 grams of s-04 and 3 grams of 71B and I did not rehydrate. I have a small anvil pump and the simple act of pumping it into the fermenter gets it nice and frothy, thats all I really do to oxygenate. I fill 1/3 of my fermenter with wort then pitch in all of the dry yeast, then fill the remaining 2/3 right on top of it to mix it in. 12 hours post pitch and it was chugging away.
Thanks! so this finished at 1.014 which is where mine typically end (or my target anyways). Im not too experienced with pitch rates, but do you have an idea of what the actual pitch rate was for this? It started at 1.072 and at 5.5gallons brewers friend shows me that 10grams total of dry yeast would yield a pitch rate of 0.27M cells/ml/P. This assumes a cell density of 10billion per gram. does this sound right? seems very low unless I of course screwed up on the inputs lol. Obviously the yeast did its job well with your beer so Im not second guessing it, but just trying to figure out what pitch rate this equated to. FWIW, your experience has been looking to purchase a packet or two of 71B lol.
 

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Thanks! so this finished at 1.014 which is where mine typically end (or my target anyways). Im not too experienced with pitch rates, but do you have an idea of what the actual pitch rate was for this? It started at 1.072 and at 5.5gallons brewers friend shows me that 10grams total of dry yeast would yield a pitch rate of 0.27M cells/ml/P. This assumes a cell density of 10billion per gram. does this sound right? seems very low unless I of course screwed up on the inputs lol. Obviously the yeast did its job well with your beer so Im not second guessing it, but just trying to figure out what pitch rate this equated to. FWIW, your experience has been looking to purchase a packet or two of 71B lol.
No idea on the pitch rate to be honest, but someone else will prob know better. Whenever using most dry yeasts I’ve always felt that one pack gets the job done for approximately 1.070ish and lower. I’ve seem some people say that nate insists on under pitching s-04 so thats kind of what I went for (7 grams) with the addition of the 71B to get to 10 grams total.
 

beervoid

Hophead & Pellet Rubber
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View attachment 685299So I carbed it up and this beer is SOFT and THICK on the palette. It still has that very fruity wine like thing going on but it’s very enjoyable to drink. My best guess is that the 71B is really contributing to the mouthfeel due to the high levels of glycerol it produces. I think I’ll try playing with some different combos going forward...both yeast combos and ratios. My next brew is going to be with omegas version of 1318 so maybe I’ll try that whole pack and 2 or 3 grams of 71B.
What grainbill did you use? And was it really much different in terms of mouthfeel with your previous batches? I did not notice any mouthfeel improvement and always do split batches to confim.
Perhaps I might have to pitch more next time.
 
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