How much total RNA is needed to isolate and clone small RNA? - (Jul/07/2008 )
Dear all,
I am using Ambion's mirVana microRNA isolation kit to enrich my small RNAs from total RNAs. This kit require 50-100 ug of total RNA in order to enrich the small RNAs. I used 50 ug of total RNAs and successful enrich the small RNAs. However, i have a lot of problems in RT-PCR. I am thinking of this is due to my starting RNAs is not enough. This is because based on some journals, they used 500 ug of total RNA to isolate small RNAs.
How do you think??
-Yew-
QUOTE (Yew @ Jul 7 2008, 05:57 AM)
Dear all,
I am using Ambion's mirVana microRNA isolation kit to enrich my small RNAs from total RNAs. This kit require 50-100 ug of total RNA in order to enrich the small RNAs. I used 50 ug of total RNAs and successful enrich the small RNAs. However, i have a lot of problems in RT-PCR. I am thinking of this is due to my starting RNAs is not enough. This is because based on some journals, they used 500 ug of total RNA to isolate small RNAs.
How do you think??
I am using Ambion's mirVana microRNA isolation kit to enrich my small RNAs from total RNAs. This kit require 50-100 ug of total RNA in order to enrich the small RNAs. I used 50 ug of total RNAs and successful enrich the small RNAs. However, i have a lot of problems in RT-PCR. I am thinking of this is due to my starting RNAs is not enough. This is because based on some journals, they used 500 ug of total RNA to isolate small RNAs.
How do you think??
Check these links for some tips on small RNA sample prep. Try out microRNA etc keywords too.
http://search.vadlo.com/b/q?sn=158621799&a...l+RNA&rel=0
hth/
-cellcounter-