RNA isolaton for microRNA profiling: Total RNA or enrichment

[zammy]

Hello everyone!!
This is my first post here and I hope it is in right section...
I have to isolate RNA from tissue samples and purpose is to perform microRNA profiling via next generation sequencing. As the first step is RNA isolation and I have Ambion mirvana kit for this. The kit has two protocols, one is Total RNA isolation and other is small RNA enrichment. I would like to know that for miRNA profiling should I follow protocol of Total RNA isolation or the small RNA enrichment protocol?

Regards
Azeem

[lrichter]

Hi Zammy;

According to the kit instructions you should use the total RNA isolation if you are doing miRNa expression profiling using miRNA arrays. From the manual: "To isolate RNA for miRNA expression profiling using miRNA arrays, we recommend following the procedure for total RNA isolation (not the enrichment procedure for small RNAs). This makes it possible to both quantitate the RNA and to critically evaluate its quality, to verify that it is suitable for array analysis. Then further purify the miRNA population (e.g., with the flashPAGE™ system) before labeling samples for miRNA array analysis."

I would think the principle would hold true for NexGen as well. Good luck.