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Author Topic: Reusable Borosilicate Screw Cap Glass Culture Tubes  (Read 12114 times)

Offline erockrph

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Re: Reusable Borosilicate Screw Cap Glass Culture Tubes
« Reply #45 on: October 30, 2015, 08:16:28 am »


If the colonies are hemispherical and smooth, you received a bonus yeast culture.  If the colonies are fuzzy, they are more than likely mold.  Personally, I would take one of the well-isolated colonies, and grow it into a pitchable culture using aseptic technique to see what you received.  However, I am one of those people who is willing to spend good money on an unknown culture, and then pitch it into wort that took several hours to prepare.  No guts, no glory! :)

No fuzz. They look like shiny/waxy white pellets. I'm brewing today, so I'll have to try this out while I'm at it.
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Offline toby

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Re: Reusable Borosilicate Screw Cap Glass Culture Tubes
« Reply #46 on: October 30, 2015, 08:33:03 am »
By the way, I thought that the yeastman website was only available to professional brewers.

No, you just have to set up a 'business account' with them (when you set up the account, "Individual" is an option).

Offline klickitat jim

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Re: Reusable Borosilicate Screw Cap Glass Culture Tubes
« Reply #47 on: December 18, 2015, 04:05:18 pm »
I've learned so much from this thread that my head hurts. I have my slants, plates, loops, lamp, tiny flasks, and agar in the shopping cart at Carolina.com just waiting for some mad money to take the leap.

Edit: Merry Christmas to me, yeast wrangling outfit is en route to my brewery!
« Last Edit: December 18, 2015, 05:30:49 pm by klickitat jim »

Offline Whiskers

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Re: Reusable Borosilicate Screw Cap Glass Culture Tubes
« Reply #48 on: December 18, 2015, 05:42:44 pm »
I've learned so much from this thread that my head hurts. I have my slants, plates, loops, lamp, tiny flasks, and agar in the shopping cart at Carolina.com just waiting for some mad money to take the leap.

Edit: Merry Christmas to me, yeast wrangling outfit is en route to my brewery!

Might check cynmar - they have pretty cheap lab stuff. 

Offline klickitat jim

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Re: Reusable Borosilicate Screw Cap Glass Culture Tubes
« Reply #49 on: December 18, 2015, 06:00:54 pm »
I've learned so much from this thread that my head hurts. I have my slants, plates, loops, lamp, tiny flasks, and agar in the shopping cart at Carolina.com just waiting for some mad money to take the leap.

Edit: Merry Christmas to me, yeast wrangling outfit is en route to my brewery!

Might check cynmar - they have pretty cheap lab stuff.
I'll do that, but I just placed my order at Carolina

Offline klickitat jim

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Re: Reusable Borosilicate Screw Cap Glass Culture Tubes
« Reply #50 on: December 18, 2015, 09:22:17 pm »
Mark,

I appologize if this is asked and answered, What is your opinion on topping off inoculated/incubated slants with autoclaved water or mineral oil for long term storage?

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Re: Reusable Borosilicate Screw Cap Glass Culture Tubes
« Reply #51 on: December 19, 2015, 05:52:54 pm »
I appologize if this is asked and answered, What is your opinion on topping off inoculated/incubated slants with autoclaved water or mineral oil for long term storage?

I have heard of labs in Eastern Europe using mineral oil, but one needs to use caps with PTFE-faced liners because mineral will react with a standard rubber liner.  The yeast cells will not remain on the surface of the media if one adds autoclaved distilled water, which makes subculturing a pain in the backside.   

In my humble opinion, it is better to set up a rotation where each culture is used at least once per year.   That way, the culture can be subcultured to a new blank slant while propagating a first level starter.  I kept CL-170 alive for over a decade using this management technique. CL-170 was the longest-lived culture in my old bank because I propagated a culture at least once per year (I repitch many cultures).  Any strain that is not used at least once every two years should be culled because it is wasting valuable real estate and brewing bandwidth.

If one wants to store yeast under autoclaved distilled water, one should carefully select one or more well-isolated colony-forming units from a plate for storage.   The key here is to ensure that no media is transferred to the distilled water.  There should only be yeast cells and H2O. This technique was taught by the ATCC in Rockville, Maryland in the nineties.  It was developed for use in countries where refrigeration is in short supply. I attended one of the sessions.  Over the years, the storing of well-isolated colonies under autoclaved distilled technique has erroneously morphed into rinsing and storing cropped yeast under boiled water via the home brewer equivalent of the game where people sit in a circle and pass information.  Anyone who has played this game realizes how distorted the information becomes as it traverses the circle.  This technique does not work the same way with cropped yeast.

Offline klickitat jim

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Re: Reusable Borosilicate Screw Cap Glass Culture Tubes
« Reply #52 on: December 19, 2015, 06:07:09 pm »
Excellent, thanks Mark. I think I will go with basic slanting and subcultuing on 6mo basis for now. Plating for me will be pretty rare at least until I get into trying to capture a local wild yeast. That probably wont be until I am well skilled with slanting. I might plate an occasional beer of my own just to see how many contaminants grow.

I suspect that im going to enjoy this aspect of the hobby. Thanks for all the help and advice

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Re: Reusable Borosilicate Screw Cap Glass Culture Tubes
« Reply #53 on: December 19, 2015, 06:17:36 pm »
Excellent, thanks Mark. I think I will go with basic slanting and subcultuing on 6mo basis for now. Plating for me will be pretty rare at least until I get into trying to capture a local wild yeast. That probably wont be until I am well skilled with slanting. I might plate an occasional beer of my own just to see how many contaminants grow.

I suspect that im going to enjoy this aspect of the hobby. Thanks for all the help and advice

You have to plate any liquid single-strain culture for singles before transferring it to slant.  Transferring a liquid culture to slant without plating it first is a no-no.

https://www.homebrewersassociation.org/forum/index.php?topic=24596.msg313974#msg313974

Offline klickitat jim

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Re: Reusable Borosilicate Screw Cap Glass Culture Tubes
« Reply #54 on: December 19, 2015, 06:27:18 pm »
Excellent, thanks Mark. I think I will go with basic slanting and subcultuing on 6mo basis for now. Plating for me will be pretty rare at least until I get into trying to capture a local wild yeast. That probably wont be until I am well skilled with slanting. I might plate an occasional beer of my own just to see how many contaminants grow.

I suspect that im going to enjoy this aspect of the hobby. Thanks for all the help and advice

You have to plate any liquid single-strain culture for singles before transferring it to slant.  Transferring a liquid culture to slant without plating it first is a no-no.

https://www.homebrewersassociation.org/forum/index.php?topic=24596.msg313974#msg313974
Awe yes, what about when subcultuing? Just loop a sample and inoculate the new slant? Then loop another to inoculate the starter?

Edit: never mind, I recall you saying to start with a single colony.

Its starting to make sense. At every step that you can, you are getting back to as pure of a pitch as you can.

From liquid sample, plate and inoculate two slants with a single colony

From the best of the two slants, plate and inoculate the starter with a single colony AND inoculate two new slants with single colonies.

Obviously using aseptic techniques betweem each step. That part I understand because of my profession. Universal precautions and Locard's Exchange Principle.
« Last Edit: December 19, 2015, 06:58:04 pm by klickitat jim »

Offline klickitat jim

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Re: Reusable Borosilicate Screw Cap Glass Culture Tubes
« Reply #55 on: December 20, 2015, 01:19:15 am »
I'm cracking myself up.

So here I am watching a YouTube video of some guy. He says "So now im going to show you how to make the slants by letting them solidify at an angle". At which point he opens the pressure cooker lid... I'm like This guy doesn't know what he's doing, he autoclaved his plates in the pressure cooker! Hey, he just touched that plate with his fingers! Hey, he doesn't have an alcohol lamp going either.

Suddenly im the expert, LOL

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Re: Reusable Borosilicate Screw Cap Glass Culture Tubes
« Reply #56 on: December 20, 2015, 08:37:26 am »
The lamp is used to create an updraft.  The updraft helps to prevent dust from settling on the media while the lid is up for pouring.  This technique does not apply to autoclaved plates because the dish is covered when the media is being sterilized.  However, anyone who has worked with plates that were autoclaved in a pressure cooker instead of using the standard laboratory technique knows that the amount of condensation that forms inside of the dish is a major problem.

Here a couple of videos of how plates are poured in a laboratory without the aid of a laminar flow hood:

https://www.youtube.com/watch?v=q7I5YN571kw

https://www.youtube.com/watch?v=gxamN2CpmX4


Two videos that demonstrate the entire process, including media preparation

https://www.youtube.com/watch?v=cneascR3OEc

https://www.youtube.com/watch?v=OljTdYH_Wtg

Did you notice the use of a magnetic stirrer?  That's what magnetic stirrers were designed to do in a laboratory.  Stir plates are for mixing, not culturing.  Like storing rinsed cropped yeast under water, the almost universal use of stir plates in home brewing for culture propagation is yet another laboratory technique/device that has been introduced via the home brewing equivalent of the pass information around a circle game. 

LB stands for Luria broth.  It is a specific type of medium.  I currently use MYGP, which stands for Malt extract, Yeast Extract, Glucose (dextrose), and Peptone (I use soytone).   However, I used agar solidified wort for many years. One does not need to use a ton of malt extract to make malt agar plates.  Two percent weight-by-volume  (w/v) is enough malt extract.  A 2% w/v malt extract solution contains 2 grams of malt extract per 100ml of solution.  Media is solidified via the addition 1.5% w/v powdered agar (Green BioResearch on eBay sells 100 grams of lab grade agar powder for a reasonable price).  In layman's terms, 1.5% w/v is 1.5 grams of powdered agar per 100ml of solution.

Remember, the updraft zone around an alcohol lamp is smaller than that of a Bunsen burner; therefore, the plates need to be closer to the heat source while pouring.  I would not pour more than 5 to 6 plates at a time unless there are many cultures to plate.

The video linked below demonstrates several basic aseptic transfer skills, including the 4-quadrant streak.  The 4-quadrant streak is one of the most fundamental skills in microbiology.  What it does dilute the culture to the point where single cells are separated on the plate.  In the case of yeast, a round, dome-shaped, opaque, white colored, non-fuzzy colony can be assumed to be the offspring of a single yeast cell; therefore, it is a pure culture.  Please note that the loop is flamed and the plate is turned 90 degrees between streaks.

https://www.youtube.com/watch?v=NV0TlN0DpPw

Finally, here is a video made by a home brewer in who is also a Ph.D. microbiologist.  I exchanged a few e-mail messages with him a year or so ago because he has access to -80C refrigeration.  He uses the old pipette a drop of liquid culture onto a piece of blotter paper that was autoclaved in aluminum foil technique to make cultures that can be mailed to other brewers in the Canadian equivalent of a first class envelope.  The blotter paper is dropped into small amount autoclaved 5% w/v wort to revive the culture and then plated for singles.  Once again, I cannot overemphasize that plating is a fundamental skill when working with yeast.

https://www.youtube.com/watch?v=-EKb6AcaEBg
« Last Edit: December 20, 2015, 05:55:21 pm by S. cerevisiae »

Offline klickitat jim

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Re: Reusable Borosilicate Screw Cap Glass Culture Tubes
« Reply #57 on: December 20, 2015, 01:34:49 pm »
Awesome! I now have a use for my stirplates again! Woo hoo

That was a great post Mark. Hopefully a lot of folks find it and remove the mystery

Offline narcout

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Re: Reusable Borosilicate Screw Cap Glass Culture Tubes
« Reply #58 on: December 20, 2015, 05:33:50 pm »
Like storing rinsed cropped yeast under water, the almost universal use of stir plates in home brewing for culture propagation is yet another laboratory technique/device that has been introduced via the home brewing equivalent of the pass information around a circle game.

White Labs staff themselves advocate the use of stir plates. It may or may not be misguided (or even bad) advice, but it doesn't seem to be the result of homebrewers playing telephone.
« Last Edit: December 20, 2015, 05:36:23 pm by narcout »
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Re: Reusable Borosilicate Screw Cap Glass Culture Tubes
« Reply #59 on: December 20, 2015, 06:11:02 pm »
White Labs staff themselves advocate the use of stir plates. It may or may not be misguided (or even bad) advice, but it doesn't seem to be the result of homebrewers playing telephone.

They clearly do so because so because of the ubiquitous use of stir plates in home brewing.  White Labs uses orbital shakers in the culture propagation lab.  Neva Parker was taken back when a member of this forum questioned the need for stirring when most brewing strains exhibit NewFlo flocculation (a.k.a. NewFlo phenotype) in the Reddit thread.

The use of stir plates in home brewing is absolutely the result of the device being introduced by a well-meaning home brewer followed by folklore.  Almost no one questions why a home brewer would actually need to use a stir plate for yeast propagation, or if the benefits outweigh the costs.