Protocol Online logo
Top : Forum Archives: : Tissue and Cell Culture

precipitation in DMEM - (Dec/19/2008 )

hey. m using powder DMEM to make medium in my lab.. We usually prepare abt 500ml in a go. after we prepare it, we store the bottles at -20 until further use. the problem is after thawing out, some of the DMEM/sodium bicarb (whatever) precipitates out n forms a layer at the base of the bottle.. we tried keeping the medium at 37 deg for thawing, hoping tht the pwdr will dissolve, but 2 no avail.
HELP pls... lot of my TC work is hanging due to this...

thanks a lot!

-Ven-

QUOTE (Ven @ Dec 19 2008, 02:23 PM)
hey. m using powder DMEM to make medium in my lab.. We usually prepare abt 500ml in a go. after we prepare it, we store the bottles at -20 until further use. the problem is after thawing out, some of the DMEM/sodium bicarb (whatever) precipitates out n forms a layer at the base of the bottle.. we tried keeping the medium at 37 deg for thawing, hoping tht the pwdr will dissolve, but 2 no avail.
HELP pls... lot of my TC work is hanging due to this...

thanks a lot!



Hi!

You can perfectly store the medium in -4 (refrigerator) with no damage!!
Here we are not used to freeze medium, cuz freeze and thaw many times can be bad.
My suggestion: prepare the medium, filter, and add suplementation ( serum, L-glutamine and antibiotics), then separate them in two aliquots ( 250 ml each) one you use daily other, store it. have no problem with this way!

Regards!

Reis, V.P.

-Reis V.P-

QUOTE (Reis V.P @ Dec 19 2008, 03:46 PM)
QUOTE (Ven @ Dec 19 2008, 02:23 PM)
hey. m using powder DMEM to make medium in my lab.. We usually prepare abt 500ml in a go. after we prepare it, we store the bottles at -20 until further use. the problem is after thawing out, some of the DMEM/sodium bicarb (whatever) precipitates out n forms a layer at the base of the bottle.. we tried keeping the medium at 37 deg for thawing, hoping tht the pwdr will dissolve, but 2 no avail.
HELP pls... lot of my TC work is hanging due to this...

thanks a lot!



Hi!

You can perfectly store the medium in -4 (refrigerator) with no damage!!
Here we are not used to freeze medium, cuz freeze and thaw many times can be bad.
My suggestion: prepare the medium, filter, and add suplementation ( serum, L-glutamine and antibiotics), then separate them in two aliquots ( 250 ml each) one you use daily other, store it. have no problem with this way!

Regards!

Reis, V.P.

-Ven-

QUOTE (Reis V.P @ Dec 19 2008, 03:46 PM)
Hi!

You can perfectly store the medium in -4 (refrigerator) with no damage!!
Here we are not used to freeze medium, cuz freeze and thaw many times can be bad.
My suggestion: prepare the medium, filter, and add suplementation ( serum, L-glutamine and antibiotics), then separate them in two aliquots ( 250 ml each) one you use daily other, store it. have no problem with this way!

Regards!

Reis, V.P.

Unless you are using the medium quite quickly, I would add the l-glutamine just before you go to use the aliquot as it decays quite rapidly in solution, somewhere around the 7-10 day mark is normal for 50% loss in medium.

-bob1-