4 replies to this topic

[mrvica]

Hi,
I need a help from you, guys. Can I save my cells (imoratlized cell line) in RNAlater for a period of time? At least a few weaks untill the TRIzol I have ordered comes? And is there any special protocol, on how to preserve cells? I`m planning to wash them three times in PBS and then to resuspend pelet in, I don`t know, maybe 200 microliters of RNAlater, and then keep the cells at -20 degrees, until RNA extraction? What do you think, will it be ok?

Thanks so much!

[bob1]

If it is just cells, why not just grow some more when the trizol arrives?

The RNAlater website advertises storage of pelleted cells as being fine.

[zodiac1505]

I have used RNAlater to save organs and usually they are fine for extended periods of time in -20. But I have always used RNAeasy kit to extract the RNA and never trizol unless the cells and organs are fresh.

[Curtis]

Do not keep at -20C. Ambion does not recommend it. keep at 4C. RNAlater is made by a guy from Ambion and he told us to keep at 4C.

also, as Zodiac suggested, if you want pure RNA for sensitive experiments (e.g Real-Time PCR) use RNeasy kit, though it has a longer protocol than a normal Trizol method.

[saraarasus]

I have used RNA later for doing experinment later. It will work. Just wash your cells and add RNA later. It should cover the cells completely. Then store it at -80 for long storage (months) or -20 for short (1-2 weeks).  

mrvica, on May 19 2010, 12:56 AM, said:

Hi,
I need a help from you, guys. Can I save my cells (imoratlized cell line) in RNAlater for a period of time? At least a few weaks untill the TRIzol I have ordered comes? And is there any special protocol, on how to preserve cells? I`m planning to wash them three times in PBS and then to resuspend pelet in, I don`t know, maybe 200 microliters of RNAlater, and then keep the cells at -20 degrees, until RNA extraction? What do you think, will it be ok?

Thanks so much!