Making Agar Slants (Slopes) - In Pictures

Share your experiences of using brewing yeast.
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greenxpaddy

Re: Making Agar Slants (Slopes) - In Pictures

Post by greenxpaddy » Wed Dec 21, 2011 4:09 pm

Image

I am looking to lift three or four colonies from the bottom of the pic. You can see where the first lines ripped the wort agar. I did go back a further two times as I rotated the plate but maybe the loop hadn't cooled right down and I forgot to cool it on the agar surface who knows.

Still seem to have some isolated colonies. Will 3 or 4 colonies be sufficient for 10ml sterilised wort?

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gregorach
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Re: Making Agar Slants (Slopes) - In Pictures

Post by gregorach » Wed Dec 21, 2011 4:20 pm

Yeah, you'll get away with that.

Streaking plates is one of those things that sounds simple enough (and in principle, it is simple enough), but really takes quite a lot of practice to get good at.
Cheers

Dunc

greenxpaddy

Re: Making Agar Slants (Slopes) - In Pictures

Post by greenxpaddy » Wed Dec 21, 2011 4:38 pm

The yeast slant was very milky, so I wondered if it was infected. You can see where it dug into the agar that it is very milky looking.

I am glad I performed this exercise because the colonies selected look really 'normal yeast'-like.

Well time will tell if the aroma is right.

Cazamodo

Re: Making Agar Slants (Slopes) - In Pictures

Post by Cazamodo » Tue Feb 07, 2012 2:21 am

How long will a blank slant last?
I made up a batch and planned on innoculating them a week later. Then I ended up not brewing, and opened my brew cupboard a month later and remembered I had a bunch in there! They dont look mouldy, just normal. So im guessing my sterilisation was better than I thought! Should I put these in teh fridge? Or just leave them untill I need to use them?

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Re: Making Agar Slants (Slopes) - In Pictures

Post by gregorach » Tue Feb 07, 2012 10:47 am

They should last more or less indefinitely in theory, as long as they're sterile and well sealed. I generally prefer to store them in the fridge, but it's not a big deal either way for blanks.
Cheers

Dunc

Cazamodo

Re: Making Agar Slants (Slopes) - In Pictures

Post by Cazamodo » Thu Mar 01, 2012 12:32 am

Cheers, after this post I then forgot about them again so they still sit in the cupboard. I was a bit dubious that they still looked good, so I opened/closed the lid of one, and this last week it has grown all kinds of gunk... So I must of had a better procedure than I thought. I assume the hard part comes keeping them sterile when inoculating.

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Re: Making Agar Slants (Slopes) - In Pictures

Post by gregorach » Thu Mar 01, 2012 10:26 am

Cazamodo wrote:I assume the hard part comes keeping them sterile when inoculating.
Indeed... The most important thing is to work next to a flame (ideally with 3 inches), and to work quickly. I tend to wear a surgical mask and gloves, and I'm pretty liberal with the hand sanitiser. The combination of nitrile gloves, alcohol gel, and an open flame does call for a great deal of caution! Plan your every move in advance - think about exactly how you're going to go about every step. Try and get into the habit of flaming the mouths of all containers as you open them and both the mouth and cap before you close them. The really tricky bit is figuring our how to open a slant and flame the mouth whilst you're still holding a loop of yeast close enough to the flame the keep it in the sterile zone, but not so close that you cook it - and all without setting yourself on fire (the trick is to unscrew the cap first, so that you can open it one-handed when you need to). I'd recommend a number of trial runs before you do it for real with a yeast that you can't easily replace.
Cheers

Dunc

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Re: Making Agar Slants (Slopes) - In Pictures

Post by jmc » Thu Mar 01, 2012 12:57 pm

gregorach wrote:
Cazamodo wrote:I assume the hard part comes keeping them sterile when inoculating.
Indeed... The most important thing is to work next to a flame (ideally with 3 inches), and to work quickly. I tend to wear a surgical mask and gloves, and I'm pretty liberal with the hand sanitiser. The combination of nitrile gloves, alcohol gel, and an open flame does call for a great deal of caution! Plan your every move in advance - think about exactly how you're going to go about every step. Try and get into the habit of flaming the mouths of all containers as you open them and both the mouth and cap before you close them. The really tricky bit is figuring our how to open a slant and flame the mouth whilst you're still holding a loop of yeast close enough to the flame the keep it in the sterile zone, but not so close that you cook it - and all without setting yourself on fire (the trick is to unscrew the cap first, so that you can open it one-handed when you need to). I'd recommend a number of trial runs before you do it for real with a yeast that you can't easily replace.
Can you use a slant as a yeast source multiple times, or is infection while taking a sample (to step up) too big a risk?

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Re: Making Agar Slants (Slopes) - In Pictures

Post by gregorach » Thu Mar 01, 2012 1:52 pm

jmc wrote:Can you use a slant as a yeast source multiple times, or is infection while taking a sample (to step up) too big a risk?
Yeah, the whole idea of slants is to be able to use them multiple times. What I like to do is inoculate at least two slants for every strain, one of which I'll use as a "working" slant, while the other remains untouched as a backup. I use different coloured labels on them so I know which is which.
Cheers

Dunc

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smeggedup
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Re: Making Agar Slants (Slopes) - In Pictures

Post by smeggedup » Sat Mar 24, 2012 7:46 pm

right
got my test tube things, agar, dme, pressure cooker, and syringe, and figure i want about 10mls for each slant, ten slants so thats 100mls of final product.

so ?
how much do i need to use of agar, water, dme ?
to get the right ratios


please would like to do these tonight

could you show working outs plz
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Re: Making Agar Slants (Slopes) - In Pictures

Post by Twistedfinger » Sat Mar 24, 2012 8:10 pm

I'd go 150ml so you have enough.

So 10% of 150 for your wort = 15g of DME to 150ml water. (1.030)
To this add about 1.5% which = 2.25g

I did some this week and added 3g of agar to 150ml by mistake and they were ok.

I've PDF'd Wolfy's instructions to print out but can't attach them here. PM me an email address and I'll send them to you if you want.
Last edited by Twistedfinger on Sat Mar 24, 2012 8:13 pm, edited 1 time in total.

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Re: Making Agar Slants (Slopes) - In Pictures

Post by jmc » Sat Mar 24, 2012 8:11 pm

I use 10% DME and 1% agar, so for 100g wort I'd use 10g Dme & 1g agar

**** edit . Pipped to the post by twistedfinger

weiht

Re: Making Agar Slants (Slopes) - In Pictures

Post by weiht » Wed Mar 28, 2012 4:43 pm

So my friend transfer some pacman yeast to my slant, and it took off really quick within 24 hrs. So, after a few days, I decided to make a starter for the upcoming brew and at the same time make another slant.

Is this normal?? infection or just the yeast going crazy?? It's actually bubble like and grows bigger, and im guessing its putting out CO2.

Image

Anyone? Dunc?

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Re: Making Agar Slants (Slopes) - In Pictures

Post by gregorach » Wed Mar 28, 2012 5:24 pm

No, that doesn't look right at all. I'd assume contamination.
Cheers

Dunc

weiht

Re: Making Agar Slants (Slopes) - In Pictures

Post by weiht » Wed Mar 28, 2012 5:28 pm

yeah, the other slants were ok except this... This slant is now having a mini fermentation inside with so much bubble and gas build up...

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