4 replies to this topic

[MrMopster]

Hello all,

I am about to start trying to do analysis of mir34a expression in primary tumour cells by Q-PCR and I need to find a cell line to be a known positive sample. I guess it needs to either over express mir34a or is guaranteed to express high levels in response to etoposide, for instance.....  Does any one have any good ideas about how to go about this? Sorry if this is a really daft question... thanks in advance for any help  

[Functional Screens]

I will suggest google "lenti mirna" or "lenti nirna expression vector" to find a virus/vector/way to make your stable cell line expressing mir-34a as positive control.

[MrMopster]

Hi Functional Screens,
Thank you for your reply, I have followed your google and this may be an option.
However, if I can find journal articles that show in vitro upregulation of this miRNA in response to drug treatment could I not use treated cells as a positive control? Or is this a bit too unreliable / wishy washy? (excuse the unscientific english). Hmm. I am hoping to get this moving quite quickly, which I could do with drug treatments but not with transfections.
Thanks again for you response  

[Functional Screens]

MrMopster, on Jun 15 2009, 08:09 AM, said:

Hi Functional Screens,
Thank you for your reply, I have followed your google and this may be an option.
However, if I can find journal articles that show in vitro upregulation of this miRNA in response to drug treatment could I not use treated cells as a positive control? Or is this a bit too unreliable / wishy washy? (excuse the unscientific english). Hmm. I am hoping to get this moving quite quickly, which I could do with drug treatments but not with transfections.
Thanks again for you response  

Hi MrMopster,
If you are just looking for mir-34a positive control for qRT-PCR, then I will suggest you to make a stable line other than spending a lot of time to seek a specific cell line which will response to an inducer.

Edited by Functional Screens, 15 June 2009 - 03:56 PM.

[MrMopster]

Functional Screens, on Jun 16 2009, 12:55 AM, said:

MrMopster, on Jun 15 2009, 08:09 AM, said:

Hi Functional Screens,
Thank you for your reply, I have followed your google and this may be an option.
However, if I can find journal articles that show in vitro upregulation of this miRNA in response to drug treatment could I not use treated cells as a positive control? Or is this a bit too unreliable / wishy washy? (excuse the unscientific english). Hmm. I am hoping to get this moving quite quickly, which I could do with drug treatments but not with transfections.
Thanks again for you response  

Hi MrMopster,
If you are just looking for mir-34a positive control for qRT-PCR, then I will suggest you to make a stable line other than spending a lot of time to seek a specific cell line which will response to an inducer.

Thanks for the advice F. S. I shall give it a try