Bacterial Stock - Protein and Proteomics

Bacterial Stock - (Sep/23/2007 )

Hi,

Can anyone tell me how to prepare bacterial glycerol stock ? What should be the perentage of glycerol?

Thanks

-veerubiotech-

QUOTE (veerubiotech @ Sep 24 2007, 01:23 PM)

Hi,

Can anyone tell me how to prepare bacterial glycerol stock ? What should be the perentage of glycerol?

Thanks

I have used either 300ul of 80% glycerol (autoclaved) and 700ul of bacteria, or 500ul of 50% glycerol and 500ul of bacteria. Both work for me.

-Almasy-

Hi,

we keep them in 15% glycerol. Prepare medium containing 30% glycerol. Mix this with the same volume of bacteria.

Cheers

-zek-

QUOTE (Almasy @ Sep 23 2007, 10:50 PM)

QUOTE (veerubiotech @ Sep 24 2007, 01:23 PM)

Hi,

Can anyone tell me how to prepare bacterial glycerol stock ? What should be the perentage of glycerol?

Thanks

I have used either 300ul of 80% glycerol (autoclaved) and 700ul of bacteria, or 500ul of 50% glycerol and 500ul of bacteria. Both work for me.

Thanks for your reply. Do we need to thaw (if yes how? )the cells before use or just scrap little bit and inoculate. Can we use again and again ? is cells wiil be viable after repeated cycle of thawing and freezing.

thanks

-veerubiotech-

QUOTE (veerubiotech @ Sep 24 2007, 02:56 PM)

QUOTE (Almasy @ Sep 23 2007, 10:50 PM)

QUOTE (veerubiotech @ Sep 24 2007, 01:23 PM)

Hi,

Can anyone tell me how to prepare bacterial glycerol stock ? What should be the perentage of glycerol?

Thanks

I have used either 300ul of 80% glycerol (autoclaved) and 700ul of bacteria, or 500ul of 50% glycerol and 500ul of bacteria. Both work for me.

No, I don't thaw it, at least not completely. What I did is that before I froze them down I mixed them (bacteria and glycerol) thoroughly. Then when I need to use, I just use the white loop, poke it inside, maybe scrape a little bit, then plate on LB agar (with antibiotic if necessary) and that is that.

-Almasy-