-20C Glycerol stock

[Sebastian Cocioba]

There are tons of different glycerol stock recipes for deep freezers but none for a standard freezer. I'm guessing a refresh rate of 6 months may be in order but what glycerol concentration? Are there any starvation issues since there will be some metabolic activity in a very limited nutrient environment. Anyone have experience with freezer stocks? Thanks.

PS I may have asked a similar question before. If so my apologies.

Sent from my Windows Phone

[Jeswin]

What are you storing? I have stored bacterial colonies using 300uL
glycerol and 200ul culture.

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[Sebastian Cocioba]

Email client sucks with replies. Sorry. Either way do you think a
simple survey of HB101 E Coli would be fine for an initial experiment
or should I pile on the different strains and species all at once? I'm
thinking 10 repeats of each concentration. Culture taken from one
colony grown up to a standard 10^8 colony forming units per mL. Streaks
made every two weeks for one year. Any ideas for better experimental
design? Thanks!

Sent from my Windows Phone From: Jeswin
Sent: 8/8/2013 4:48 PM
To: Sebastian Cocioba
Subject: Re: [DIYbio] -20C Glycerol stock
On Thu, Aug 8, 2013 at 4:34 PM, Sebastian Cocioba <scoc...@gmail.com> wrote:
> Agro, ecoli, etc...usual stuff. Just curious if anyone has any hard
> data on the effects of varying gycerol concentrations on viability long
> term. If not then that sounds like a simple passive experiment to run.
>

You should have replied to the list.

Anyway, if you're going to do an experiment, it would be nice to know.

[SC]

Hi Sebastian,

 

I use one part bacterial culture and one part sterile glycerol.  Mix thouroughly with a wide bore pipette (or chop the end off a 1000 ml blue tip.)  They last at least a year at -20oC, maybe longer. (E coli-based clones).

 

Note:  An agar stab in an eppy with a gas permeable top and stored in the fridge at 4oC lasts a few months.  I use that method when I have a larger queue of clones than I can deal with right away but don't need long term storage.  Hint:  Gas permeable bandages with sticky on the back make a great topper for these. Just cut it up, stick on, and fold the edges over.   If you have a lot of clones use a 96 well plate and a whole bandage.

 

Good luck!

 

Stacy

 

[Sebastian Cocioba]

Thanks for the info, Stacy! Neat trick with the gas permeable tape. I'll definitely look into that.

Sent from my Windows Phone

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From: SC
Sent: 8/12/2013 6:46 PM
To: diy...@googlegroups.com
Subject: [DIYbio] Re: -20C Glycerol stock

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