Gene silencing and miRNA isolation - (Apr/26/2014 )

Hi everyone,

In my project, I will first try to silence a gene and then will look for its effects on miRNAs in cells(eukaryotic). 

Basically, i will use lipofectamine reagent and specific siRNA to silence the gene(say X gene).

1) To show that silencing is accomplished, I will isolate total RNA, convert to cDNA and run a qPCR reaction. Then,

2) I will analyse miRNAs in the X gene-silenced cells. For that, I need to isolate miRNAs and send them to RNA sequencing.

So my question is, how can I do 1 and 2 using the same sample? 

I need to do them using the same sample, because i need to look for specific miRNAs in the X gene-silenced cells. Same sample or another possible way, I am waiting to hear your suggestions. 

Thanks in advance.

Regards,

Fatih

Hi,

You can isolate total RNA include miRNAs to use Trizol reagent, so you can also obtain miRNAs