3 replies to this topic

[Aideen]

Hi,

Ive just opened this account today, Im new at this so please be patient! Ive been reading through some previous posts regarding the extraction of miRNAs using the trizol method, I cant actually find how to get back to that page or if it exists anymore but if anyone could help me I would really appreciate it!

Ok so what Im wondering is.....I want to use the Trizol method to extract microRNAs, should I follow the original protocol for extraction of total RNA or do I need to modify the Isopropanol precipiation step and also the ethanol wash??

Aideen

[Clare]

Welcome to Bioforum

I am sure you'll find loads of helpful advice from friendly people!

For Trizol and miRNAs we just use the same method. Our first lot of results from some microRNA expression arrays will be coming back soon! So YAY! Will keep you posted!
Clare

Aideen, on Aug 25 2009, 11:03 AM, said:

Hi,

Ive just opened this account today, Im new at this so please be patient! Ive been reading through some previous posts regarding the extraction of miRNAs using the trizol method, I cant actually find how to get back to that page or if it exists anymore but if anyone could help me I would really appreciate it!

Ok so what Im wondering is.....I want to use the Trizol method to extract microRNAs, should I follow the original protocol for extraction of total RNA or do I need to modify the Isopropanol precipiation step and also the ethanol wash??

Aideen

[Aideen]

[quote name='Clare' date='Aug 25 2009, 12:57 PM' post='34316']
Welcome to Bioforum

I am sure you'll find loads of helpful advice from friendly people!

For Trizol and miRNAs we just use the same method. Our first lot of results from some microRNA expression arrays will be coming back soon! So YAY! Will keep you posted!
Clare

[quote name='Aideen' post='34300' date='Aug 25 2009, 11:03 AM']Hi,

Ive just opened this account today, Im new at this so please be patient! Ive been reading through some previous posts regarding the extraction of miRNAs using the trizol method, I cant actually find how to get back to that page or if it exists anymore but if anyone could help me I would really appreciate it!

Ok so what Im wondering is.....I want to use the Trizol method to extract microRNAs, should I follow the original protocol for extraction of total RNA or do I need to modify the Isopropanol precipiation step and also the ethanol wash??

Aideen


Thanks alot Clare!

[Biodate]

For microRNA extraction and total RNA, we use a kit from Norgen Biotek. Total RNA without phenol and I don't enrich for miRNA, but I did once and used a second column to fractionate the small from the large using the same kit.

Trizol works, but not when I have small sample input. I find that the alcohol ppt step gives variable results.

Cheers