Log in


Home  |  Protocols  |  Molecular Biology  |  RNA riboprobe synthesis
Home  |  Protocols  |  Molecular Biology  |  RNA riboprobe synthesis

Protocol - RNA riboprobe synthesis

T7 RNA polymerase transcription

CategoryMolecular Biology
Last revisionMay 22, 2015
Author(s)Laura Pisapia, Pasquale Barba, Giovanna Del Pozzo
NameGiovnna Del Pozzo
AddressIGB- CNR


Description Temperature Time Note
Prepare DNA template by PCR
3-4 hrs
Use 0,5 ug of template to perform T7 RNA in vitro transcription by MaxiScript T7 kit ( AMBION) in presence of 40 uCi 32P UTP
1,5 hrs
Purify the RNA labelled riboprobe by micro Bio Spin Column P30 Tris RNase free ( BIORAD)
15 min
Measure 32UTP incorporation by beta-counter
30 min


Quality validation: Yes

Validation info

The protocol has been used to produce data published in peer-reviewed journals.


Pisapia L, Cicatiello V, Barba P, Malanga D, Maffei A, Hamilton RS, et al. Co-regulated expression of alpha and beta mRNAs encoding HLA-DR surface heterodimers is mediated by the MHCII RNA operon. Nucleic Acids Res 2013;41:3772-3786.
Corso C, Pisapia L, Citro A, Cicatiello V, Barba P, Cigliano L, et al. EBP1 and DRBP76/NF90 binding proteins are included in the major histocompatibility complex class II RNA operon. Nucleic Acids Res 2011;39:7263-7275.


Document Actions
by Abstract