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looking for a protocol for in vitro treatment of tissues - (Oct/30/2006 )

Hi,

I would like to study the effects on gene expression of some endocrine disruptors in tissues but I have no idea how can I make it. I already have the frozen tissues and my idea was to treat [i]in vitro the tissues and then extract the RNA. I need help with the buffer I should use and the best protocol to obtain the RNA.
I hope you can help me. Thank you

-Carol06-

QUOTE (Carol06 @ Oct 30 2006, 04:50 PM)
Hi,

I would like to study the effects on gene expression of some endocrine disruptors in tissues but I have no idea how can I make it. I already have the frozen tissues and my idea was to treat [i]in vitro the tissues and then extract the RNA. I need help with the buffer I should use and the best protocol to obtain the RNA.
I hope you can help me. Thank you


what do you mean by "treat in vitro"? do you mean functional assays and then extracting RNA?

-The Bearer-

That’s right. I would like to add my chemicals to my homogenized samples and after that extracting the RNA and study expression of estrogen-responsive genes. I’m not sure if this is the best method but I had liver samples and I think that could be a possibility. Can anyone suggest any other possible methods? Someone working on something related?

Thanks

-Carol06-