To label newly synthesized RNA, cells were incubated for 2 hours with [3H]-uridine (1.2 μCi/ml; PerkinElmer). Cells were counted, and the total RNA corresponding to an equal number of cells for each condition was separated by Northern blot having for the time = 0 1 μg of total RNA. Agarose-resolved RNAs were transferred to Hybond N+ membrane (GE Healthcare). After ultraviolet cross-linking, the membrane was sprayed with EN3HANCE (PerkinElmer) and exposed to Kodak BioMax MS film (Kodak) at −80°C for 1 week (22 (link)). Nascent rRNA was detected by autoradiography.
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Labeling Newly Synthesized RNA
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Corresponding Organization : Universitat de Barcelona
Other organizations : Institut Català d'Oncologia, Centro de Investigación Biomédica en Red de Epidemiología y Salud Pública, Bellvitge University Hospital, Centro de Investigación Biomédica en Red de Cáncer
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Variable analysis
independent variables
- Incubation time with [3H]-uridine (2 hours)
- Nascent rRNA detected by autoradiography
- Equal number of cells for each condition
- 1 μg of total RNA for the time = 0 condition
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