Biotinylated utps

Manufactured by Roche

Sourced in United States

Biotinylated UTPs are nucleotides that have been modified by the addition of a biotin molecule. They are used in various molecular biology and biochemical applications, such as labeling and detection of nucleic acids.

Automatically generated - may contain errors

Lab products found in correlation

Sp6 polymerase, by Roche (1 mentions) Sp6 or t7 polymerase, by Roche (1 mentions)

2 protocols using biotinylated utps

Biotinylated RNA Probe Pulldown Assay

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The pCRII-FAS-3′UTR, pCRII-HIF1A-3′UTR, pCRII-HIF1A-3′UTR-delta2D, pCRII-MTHFD2–3′UTR and pCRII-MTHFD2–3′UTR-delta2D vectors were linearized with Not1 or XhoI and used as templates for in vitro transcription using Sp6 polymerase (Roche, Indianapolis, IN, USA) in the presence of biotin labeling mix containing biotinylated UTPs (Roche) according to the manufacturer's protocol. 7 nM of probes were then incubated with approximately 500 μg of cell lysate of HCT116 cells transiently transfected with pCMVtag2bhWig-1 and used for the pulldown as described in [7 (link)].

Bersani C., Huss M., Giacomello S., Xu L.D., Bianchi J., Eriksson S., Jerhammar F., Alexeyenko A., Vilborg A., Lundeberg J., Lui W.O, & Wiman K.G. (2015). Genome-wide identification of Wig-1 mRNA targets by RIP-Seq analysis. Oncotarget, 7(2), 1895-1911.

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FAS mRNA Binding Protein Assay

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HCT116 or Saos-2 cells were harvested 24 h after transfection or 48 h after induction with 1 μg/ml doxycycline, respectively. RNA-IP was performed as described in Vilborg et al.¹⁴ For biotin pull-down assays, the pCRII-FAS-5′-UTR-ORF, pCRII-FAS-ORF-3′-UTR, pCMV6AC-FAS-distal-3′-UTR, and pCS2(+)-FAS-distal-dARE vectors were linearized with Not1 or XhoI and used as templates for in vitro transcription using T7 or Sp6 polymerase (Roche, Indianapolis, IN, USA) in the presence of biotin-labeling mix containing biotinylated UTPs (Roche) according to the manufacturer’s protocol. 10 nM probes were then incubated with approximately 200 μg of cell lysate of HCT116 p53^−/− cells transiently transfected with pCMVtag2bhWig-1 and used for the pull-down as described.^{14 (link)}

Bersani C., Xu L.D., Vilborg A., Lui W.O, & Wiman K.G. (2014). Wig-1 regulates cell cycle arrest and cell death through the p53 targets FAS and 14-3-3σ. Oncogene, 33(35), 4407-4417.

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