3' oligolabeling non radioactive end-labeling - (Aug/23/2007 )

Hello everybody,

I plan to perform Northern Blots with miRNAs from parasites.
For detection of the miRNAs (~21nt length) I'll design probes that are end-labeled.

Has anybody an idea or protocol for 3' oligolabeling non radioactive end-labeling of small RNA probes?

Thanks a lot!
-Mariana-

For better result, I recommend you ordering 5' or 3' Dig labled RNA oligo or LNA oligos, you could found from Exiqon catalog (very expensive ).
If you prefer labeling your probe in lab, you might using 3 labeling kit provided by Roche, which could label probe with ddUTP-Dig by TdT.
Another choice is labeling probe by TdT using dUTP-Dig and dATP, which generate a long tail with Dig (I have not test this idea )

Ramkissoon SH, Mainwaring LA, Sloand EM, Young NS, Kajigaya S.
Nonisotopic detection of microRNA using digoxigenin labeled RNA probes.
Mol Cell Probes. 2006 Feb;20(1):1-4.

Martin, Ruth C.; Liu, Po-Pu; Nonogaki, Hiroyuki
Simple purification of small RNAs from seeds and efficient detection of multiple microRNAs expressed in Arabidopsis thaliana and tomato (Lycopersicon esculentum) seeds
Seed Science Research, Volume 15, Number 4, December 2005 , pp. 319-328(10)