Slanting yeast

Yakima 728
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Saccharomyces said:
I jab it into the slant but not all the way to the bottom. Streaks of yeast will grow everywhere the clip has touched, each a colony from only a few cells.
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Not sure if this has been covered before. It's a big thread and I didn't read all the way through it. Apologies if this isn't new.

Stabbing the paperclip (or loop) into the media isn't necessary. All you need to do is smear the yeast on the surface.

When I started out doing this I stabbed all the way into the media. I would frequently have slants that blew out the top of the vial. Out of 8 slants that I innoculated, I would have one or two that were unusable. The yeast would ferment underneath the media and blow it out the top.

Now I just smear the yeast onto the surface. Very little yeast is required. No more stabbing means no more wasted slants and I also get more of the yeast out of the vial when the time comes to make a starter.
 
I keep having issues with some water on top of my media in the slant after the autoclave. I am using the 7/1/1.5% to 100ml (I think those are the proportions) recipe with powdered agar agar.

Any ideas?
 
Thanks for this thread! Getting yeast strains here is a huge pain in the ass and anything I can do to propagate and stretch out the useful life of what I can get here is a huge benefit to me.
 
thatjonguy said:
I keep having issues with some water on top of my media in the slant after the autoclave. I am using the 7/1/1.5% to 100ml (I think those are the proportions) recipe with powdered agar agar.

Any ideas?
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Assuming your slants aren't leaking and letting water in, this is normal. I make up a bunch at a time and store in the fridge until I need them, then take them out to warm up I put it on its top. After its warm and when I'm ready to use I open it and let the water drip out before inoculating. I haven't had any problems.
 
k_mcarthur said:
Assuming your slants aren't leaking and letting water in, this is normal. I make up a bunch at a time and store in the fridge until I need them, then take them out to warm up I put it on its top. After its warm and when I'm ready to use I open it and let the water drip out before inoculating. I haven't had any problems.
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Thanks, I heard it was normal bit wanted to find a way to correct it. I didn't want to try dumping out water without some advice.
 
MangoMead said:
Thanks for this thread! Getting yeast strains here is a huge pain in the ass and anything I can do to propagate and stretch out the useful life of what I can get here is a huge benefit to me.
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I agree. I've been washing yeast for years, finally got the stuff to plate and slant a few months ago. It takes some planning to step up from a scraping to a pitchable size, but we are capable of that. I've saved a couple hundred dollars over the last few years reusing yeast, and plating has opened up more fridge space for more beers!
 
Yeah I couldn't even tell you how much money slanting has saved me. I only buy yeast when it's a strain I don't already have.

I bought a package of Nottingham and rehydrated it for a batch I brewed. Before I pitched it, I inoculated 8 slants. So the next 8 times I need Nottingham, I just need 48 hours to build up the population.
 
I'm sure this has been answered, but I wanted to confirm. The viles that you get the White labs yeast in can be used for slanting correct? I'm sure they can handle it just want to make sure.
 
tally350z said:
I'm sure this has been answered, but I wanted to confirm. The viles that you get the White labs yeast in can be used for slanting correct? I'm sure they can handle it just want to make sure.
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You need to be able to "autoclave" them in a pressure cooker. Not sure how the plastic would hold up that. You could try soaking in star san and pour boiling hot agar solution in to help sterilize. Never used them myself but it's worth a shot, although I get Pyrex slants off Amazon for less than $10 delivered. I know they work.
 
I wouldn't use them. They're too big for one. I don't know if they would hold up inside the pressure cooker at 250* either.
 
tally350z said:
I'm sure this has been answered, but I wanted to confirm. The viles that you get the White labs yeast in can be used for slanting correct? I'm sure they can handle it just want to make sure.
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I've wondered the same thing, since I have so many hanging around, and some plastics are autoclavable, but like a previous poster said, they're pretty big. If I had a junky pressure cooker I could toss out if the vials melted, I'd experiment with them, but I'm just not gonna risk it.

Price-wise, the 24ml vials with autoclavable caps from Cynmar are 10 bucks a dozen. The hardest part for me was getting my order up to their $25 minimum. Shipping was pricey, but for 24 vials, a nichrome loop and an alcohol lamp, it was $35 including shipping. Worth not risking messing up my pressure cooker with melted vials!
 
DWhitwell said:
I've wondered the same thing, since I have so many hanging around, and some plastics are autoclavable, but like a previous poster said, they're pretty big. If I had a junky pressure cooker I could toss out if the vials melted, I'd experiment with them, but I'm just not gonna risk it.

Price-wise, the 24ml vials with autoclavable caps from Cynmar are 10 bucks a dozen. The hardest part for me was getting my order up to their $25 minimum. Shipping was pricey, but for 24 vials, a nichrome loop and an alcohol lamp, it was $35 including shipping. Worth not risking messing up my pressure cooker with melted vials!
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Very simple, just put 2 or 3 white labs vials with water into a beaker and autoclave them for the prescribed 15 mins or so and then pull them out. If they melted at all, the melted plastic will have all ended up in the beaker and you won't have ruined your autoclave. Another way to find out more about this is to look through more of this thread. From what I remember in earlier posts, White Labs uses soda preform bottles and I believe that the plastic used is rated just above 250F. Someone should still test it and post their results. I use true glass vials and don't have any white labs vials handy otherwise I would do this in a heartbeat.
 
^ That is a good test.

Yes, they are soda preforms. I've seen them for sale on Amazon.
White Labs must sterilize them too. If they don't use an autoclave in the packaging line, then what would they be using? UV? Chlorine dioxide?
 
IslandLizard said:
^ That is a good test.

Yes, they are soda preforms. I've seen them for sale on Amazon.
White Labs must sterilize them too. If they don't use an autoclave in the packaging line, then what would they be using? UV? Chlorine dioxide?
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My guess would not be chlorine dioxide because of chemical residue. UV potentially. I bet you could email them at white labs to find out. It just seems that autoclave is the preferred sterilization technique for biological assays, which is exactly what we're all basically doing when trying to wrangle and ranch yeast.
 
fc36 said:
My guess would not be chlorine dioxide because of chemical residue. UV potentially. I bet you could email them at white labs to find out. It just seems that autoclave is the preferred sterilization technique for biological assays, which is exactly what we're all basically doing when trying to wrangle and ranch yeast.
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I've seen on a television show where surgical supply manufacturers use radiation after packaging to sterilize. Maybe they radiate before packaging the yeast.
 
The agar-agar they sell at my local asian store is bright green, looks like the ingredients are agar-agar and green dye. Is this ok?
 
Since this topic has been revived, here's an issue I've been wrestling with. Perhaps someone here can offer a suggestion.

How do you get the slant out of the vial after you're done with it and want to clean and re-use the vial? I've run them through my dishwasher (which gets hot enough to sanitize things), but it still leaves chunks of agar in the vials (albeit deformed). What's the easiest way to get the spent slant completely out of the vials so they can be re-slanted?
 
kombat said:
Since this topic has been revived, here's an issue I've been wrestling with. Perhaps someone here can offer a suggestion.

How do you get the slant out of the vial after you're done with it and want to clean and re-use the vial? I've run them through my dishwasher (which gets hot enough to sanitize things), but it still leaves chunks of agar in the vials (albeit deformed). What's the easiest way to get the spent slant completely out of the vials so they can be re-slanted?
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I boil them and the agar solution becomes liquid again, use tongs to dump and rinse in the boiling water. That seems to work fine for me.
 
Hi all

I'd like to start yeast slanting, should I find any glass vials with screw caps in my s h i t'o'country.:mad:

Can Dextrose be used as for yeasts' nutrition instead of DME in the growth medium?
Glucose, Dextrose (D-Glucose) is the yeasties beasties growth nutriment of choice so I think it will work.:D

I have some Dextrose lying around and I'm not too keen on spending extra for the DME.

Greets.
 
kombat said:
How do you get the slant out of the vial after you're done with it and want to clean and re-use the vial?
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I use a long, slender kitchen tool - like a chopstick - and cut the slant up into four pieces inside the vial. One good shake over the sink and they come out pretty easily. Then I fill the vial and cap up with hot water and sprinkle in a tiny bit of oxyclean. They come out super clean.

If I have a bunch I'll put them in a glass container, fill the container all the way up to the top so the vials are all completely submerged, then put it in the microwave for a couple of minutes to boil and dissolve the agar. Then dump and rinse.

kukubau said:
Can Dextrose be used as for yeasts' nutrition instead of DME in the growth medium?
Glucose, Dextrose (D-Glucose) is the yeasties beasties growth nutriment of choice so I think it will work.:D
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I wouldn't use dextrose for the same reason I wouldn't use it in a starter. You want the yeast to acclimate themselves to metabolize maltose and the nutrients in wort. They will perform better when you ask them to metabolize something that they're used to.

I have some Dextrose lying around and I'm not too keen on spending extra for the DME.
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How are you planning on growing each slant culture up to enough yeast to pitch for a batch if you won't use malt extract?
 
LovesIPA said:
I use a long, slender kitchen tool - like a chopstick - and cut the slant up into four pieces inside the vial. One good shake over the sink and they come out pretty easily. Then I fill the vial and cap up with hot water and sprinkle in a tiny bit of oxyclean. They come out super clean.

If I have a bunch I'll put them in a glass container, fill the container all the way up to the top so the vials are all completely submerged, then put it in the microwave for a couple of minutes to boil and dissolve the agar. Then dump and rinse.

I wouldn't use dextrose for the same reason I wouldn't use it in a starter. You want the yeast to acclimate themselves to metabolize maltose and the nutrients in wort. They will perform better when you ask them to metabolize something that they're used to.

How are you planning on growing each slant culture up to enough yeast to pitch for a batch if you won't use malt extract?
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I save runnings from a mash as my starter liquid. You can always save another liter or two from a mash. Unless your system is super efficient.
 
Gothic_Horror said:
I save runnings from a mash as my starter liquid. You can always save another liter or two from a mash. Unless your system is super efficient.
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You can do that if you brew often enough, but I don't have a regular schedule. It might be 4 days before I brew again, or it might be two weeks.

It's not a good idea to leave even boiled wort around longer than a day or two. Boiling doesn't kill everything. That's why the slants are boiled in a pressure cooker.
 
LovesIPA said:
You can do that if you brew often enough, but I don't have a regular schedule. It might be 4 days before I brew again, or it might be two weeks.

It's not a good idea to leave even boiled wort around longer than a day or two. Boiling doesn't kill everything. That's why the slants are boiled in a pressure cooker.
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That's why I pressure cook and can my starter wort.
 
I can't seem to find anyone in my country to sell me glass vials in small quantities, only at least 100 pcs. and up and at an astronomic price, $400 for 100 20 ml/6 dram tubes.

I'm willing to pay you guys upfront for the glass vials and shipping, through paypal or check. Will you help me?

My only accessible alternative are the polypropylene/plastic vials, which I'm reluctant in using them.
 
kukubau said:
I can't seem to find anyone in my country to sell me glass vials in small quantities, only at least 100 pcs. and up and at an astronomic price, $400 for 100 20 ml/6 dram tubes. I'm willing to pay you guys upfront for the glass vials and shipping, through paypal or check. Will you help me? My only accessible alternative are the polypropylene/plastic vials, which I'm reluctant in using them.
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Look on eBay... That's where I found mine. ~$40 for 100 pieces as I recall. Let me know if you still can't find them and I'll get on a computer and log in to eBay to see if I can find the vendor. I can't do this on my phone.
 
kukubau said:
I can't seem to find anyone in my country to sell me glass vials in small quantities, only at least 100 pcs. and up and at an astronomic price, $400 for 100 20 ml/6 dram tubes. I'm willing to pay you guys upfront for the glass vials and shipping, through paypal or check. Will you help me? My only accessible alternative are the polypropylene/plastic vials, which I'm reluctant in using them.
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Use this service: http://www.shipito.com/?id_affiliate=2044
 
Thanks for the reply but I've already searched on ebay. No US seller will ship overseas and no European seller has the vials I need, only 2 dram which are too small. I've found a Chinese seller, but I don't think the tubes he's selling are heat resistant (I don't like the term autoclavable as my pressure cooker is nothing like an autoclave :D).

Here's the ebay bid: 10 Glass Test Tube w/ Screw Cap 15 x 150 mm - 6 Inch. 30 pcs at $42.57

The best I could found. Free shipping sounds good, but they're still too expensive for my budget.
 
k_mcarthur said:
Autoclavable is what you want. Pressure cooker is just like an autoclave.
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I know. I'm just saying I don't like the term "autoclavable" used for the pressure cooker. I used to work with autoclaves back in uni. and they're nothing alike. :)
.
 
kukubau said:
I know. I'm just saying I don't like the term "autoclavable" used for the pressure cooker. I used to work with autoclaves back in uni. and they're nothing alike. :)
.
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Sure, one is less than $100 and the other is more than $2000. I have never seen a product labeled "pressure cookerable", so we are stuck looking for "autoclavable". You may know the differences but we shouldn't confuse newer brewers reading this trying to figure this stuff out.
 
k_mcarthur said:
Sure, one is less than $100 and the other is more than $2000. I have never seen a product labeled "pressure cookerable", so we are stuck looking for "autoclavable". You may know the differences but we shouldn't confuse newer brewers reading this trying to figure this stuff out.
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We shouldn't.

I like it how "pressure cookerable" sounds. :)
 
As I understand it, pressure cookers are not quite as effective as a true pressure canner. From what I've read, pressure cookers only get up to around 12 psi. Pressure canners get up to 15 psi, which is better for truly sterilizing the contents.
 
kombat said:
As I understand it, pressure cookers are not quite as effective as a true pressure canner. From what I've read, pressure cookers only get up to around 12 psi. Pressure canners get up to 15 psi, which is better for truly sterilizing the contents.
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So I may have missed something in the many pages here, but the original post says to set the canner to 12 psi. Are you saying this won't work? It doesn't matter in my case, because my cooker specifies it gets to 15 psi, 250F, but I am still curious.
 
DWhitwell said:
So I may have missed something in the many pages here, but the original post says to set the canner to 12 psi. Are you saying this won't work? It doesn't matter in my case, because my cooker specifies it gets to 15 psi, 250F, but I am still curious.
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To be clear, I've followed this method twice, using a pressure cooker (not canner) that only gets to around 12 psi. The first time, I had about an 80% failure rate in my agar slants (that is 4 out of 5 were contaminated). I had "autoclaved" them for 15 minutes in my pressure cooker.

The second time, I autoclaved them for 25 minutes and was more careful with my handling (I wore latex gloves, dipped my hands in Star-San, screwed the caps down while the vials were still in the pressure cooker chamber, etc.), so this technique definitely can work with a simple pressure cooker, even if it only gets to 12 psi.

That said, I'm getting ready to can a batch of starter wort, and the literature I've read on canning specifies that 15 psi is necessary to ensure total sterilization. To that end, I've purchased an actual pressure canner for the task, and will follow the recommendations of 15 psi to decrease the risk of wasting my time.
 
I've stumbled upon a great website useful for the average Joe yeast grower.

http://everythingmushrooms.com

They have everything from pyrex tubes to DME and pressure cookers.

Again totally useless to me as the shipping fees to my country are $50.:mad:
 

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