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Home  |  Protocols  |  Molecular Biology  |  Magnetic labeling and isolation of biotinylated molecules
Home  |  Protocols  |  Molecular Biology  |  Magnetic labeling and isolation of biotinylated molecules

Protocol - Magnetic labeling and isolation of biotinylated molecules

A fishing/competition approach used to validate the interaction between lncRNAs and microRNAs by using biotinylated Peptide Nucleic Acid (PNA) oligomers as probes.

CategoryMolecular Biology
Creation dateOct 10, 2015
Last revisionJul 15, 2016
Author(s)Sara Terreri
NameAmelia Cimmino


Figure legend:

Expression of miR-596 in J82 cell extracts after retrieval of endogenous uc.8+ with a peptide nucleic acid (PNA)/uc.8+ probe. Data are expressed as the means ± standard deviation (SD) of triplicate values. P values were obtained using the Student t test for independent samples. ***P<0.001 (from Olivieri et al., 2016)


Description Temperature Time Note
Total RNA (300 μg) extraction
RNA incubation with 5'-biotinylated PNA oligomer
with rotation
μMACS Streptavidin MicroBeads (100 μL, Miltenyi Biotec) addition
μMACS column equilibration
100 μL of equilibration buffer for nucleic acids
RNA::PNA complex addition on the top of the μMACS column pre-equilibrated
Column washes
4x100 μL washing buffer
RNA labeled with biotynilated PNA oligomer elution
150 μL elution buffer

Other informations

100 μL μMACS™ Streptavidin MicroBeads bind up to 100 pool biotinylated molecules.


Quality validation: Yes

Validation info

The protocol has been published in a peer reviewed journal.


Olivieri M*, Ferro M*, Terreri S*, Durso M, Romanelli A, Avitabile C, De Cobelli O, Messere A, Bruzzese D, Vannini I, Marinelli L, Novellino E, Zhang W, Incoronato M, Ilardi G, Staibano S, Marra L, Franco R, Perdonà S, Terracciano D, Czerniak B, Liguori GL, Colonna V, Fabbri M, Febbraio F, Calin GA, Cimmino A. (*= equal contribution) Long non-coding RNA containing ultraconserved genomic region 8 promotes bladder cancer tumorigenesis. Oncotarget. 2016 Mar 1. doi: 10.18632/oncotarget.7833.


Funded byAIRC


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