2 replies to this topic

[valh]

Hi,
I try to detect small RNAs with a low expression level in human cells.
I want to enrich total RNA extracts in small RNAs to allow higher input levels for analysis by northern blot.
Do you know a procedure to do that ? Is the new kit "mirVana miRNA Isolation kit" from Ambion efficient ?
Thank you for help

[phdconsult]

The Ambion kit is helpful but the starting material quantities they recommend must be taken with caution. We have found the protocol to be efficient when cell batches are subdivided into smaller quantities, run the process and the final material pooled. Good luck

[flashboy]

i've just used the ambion kit to isolate small RNA's from human adipose tissue... using small chunks (i guess about 20-50mg) of tissue i averaged about 10-20ng/ul of RNA (nanodrop)