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_19
Snippet: The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/261776 doi: bioRxiv preprint pUC57-globin-Fluc was linearised with FspI and pUC57-ZIKV-Fluc was linearised with HindIII. RNA was transcribed using recombinant T7 polymerase at a final concentration of 50 ng/μL in transcription buffer (40 mM HEPES pH 7.5, 32 mM MgOAc, 40 mM DTT, 2 mM Spermidine, 10 mM NTPs, 0.2 U/μL RNaseOUT (Invitrogen)) for 2-.....
Document: The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/261776 doi: bioRxiv preprint pUC57-globin-Fluc was linearised with FspI and pUC57-ZIKV-Fluc was linearised with HindIII. RNA was transcribed using recombinant T7 polymerase at a final concentration of 50 ng/μL in transcription buffer (40 mM HEPES pH 7.5, 32 mM MgOAc, 40 mM DTT, 2 mM Spermidine, 10 mM NTPs, 0.2 U/μL RNaseOUT (Invitrogen)) for 2-4 hours at 37 C. RNA was purified by DNaseI treatment, acidic phenol extraction and ethanol precipitation. Residual nucleotides were removed using Illustra MicroSpin G-50 columns (GE Healthcare). RNA was capped using ScriptCap system and ScriptCap 2'-O-methyltransferase (CellScript).
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