Selected article for: "primer reverse and reverse primer"
Author: Renata C Fleith; Harriet V Mears; Edward Emmott; Stephen C Graham; Daniel S Mansur; Trevor R Sweeney
Title: IFIT3 and IFIT2/3 promote IFIT1-mediated translation inhibition by enhancing binding to non-self RNA
  • Document date: 2018_2_8
    • ID: j97gul0w_21
    Snippet: Analysis of the IFIT1-mRNA interaction by inhibition of primer extension (toeprinting) IFIT1/mRNA interaction was performed essentially as described previously (13) with minor modifications. Cap0-ZV or cap0--globin reporter mRNAs (1 nM) were incubated with IFIT1 or IFIT1 containing complexes (at concentrations indicated in Figures) for 10 min at 37 C in 20 μL reactions containing 20 mM Tris pH 7.5, 100 mM KCl, 2.5 mM MgCl2, 1 mM ATP, 0.2 m.....
    Document: Analysis of the IFIT1-mRNA interaction by inhibition of primer extension (toeprinting) IFIT1/mRNA interaction was performed essentially as described previously (13) with minor modifications. Cap0-ZV or cap0--globin reporter mRNAs (1 nM) were incubated with IFIT1 or IFIT1 containing complexes (at concentrations indicated in Figures) for 10 min at 37 C in 20 μL reactions containing 20 mM Tris pH 7.5, 100 mM KCl, 2.5 mM MgCl2, 1 mM ATP, 0.2 mM GTP, 1 mM DTT, 0.25 mM spermidine and 0.5 mg/mL BSA. IFIT1/mRNA interaction was monitored by inhibition of primer extension using avian myeloblastosis virus reverse transcriptase (2.5 U) (Promega) and a 32 P-labelled primer in the presence of 4 mM MgCl2 and 0.5 mM dNTPs. Full length and truncated cDNA products were separated in a denaturing 6 % acrylamide gel and detected by autoradiography using an FLA7000 Typhoon Scanner (GE). Analysis was performed using Image-Quant TL.

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