Dataset Of High-Throughput Ligand Screening Against the RNA Packaging Signals Regulating Hepatitis B Virus Nucleocapsid Formation
Raw data and average/SEM calculations for the article entitled: "Dataset Of High-Throughput Ligand Screening Against the RNA Packaging Signals Regulating Hepatitis B Virus Nucleocapsid Formation"
Steps to reproduce
NCP Assembly Assays Hepatitis B Virus (HBV) genomic RNA and PS1 RNA oligonucleotides were heat-annealed by heating to 70 oC, cooling slowly to room temperature in a buffer containing 10 mM MES pH 7.0, 25 mM NaCl and 1 mM DTT. RNA substrates were then diluted to working concentrations of 1.1 / 16.5 nM respectively in a buffer containing 25 mM HEPES pH 7.5, 250 mM NaCl and 5 mM DTT. 178 µL of RNA substrate was titrated using a Biomek 4000 liquid handling robot (Beckmann Coulter) into the wells of a 96 well plate (Greiner Bio-One, product no. 655900) and allowed to equilibrate at room temperature for 30 mins. 2 µL DMSO ± 10 µM compound 1 - 66 were added and a further equilibration step performed. Purified HBV Core Protein (Cp) dimer was then titrated using the Biomek 4000 liquid handling robot stepwise into the RNA substrates as detailed in Table 1, up to a ratio of 1:1200 (RNA:Cp dimer). 1 µM RNase A was added when assembly reactions were complete. Fluorescence anistropy was monitored throughout using a POLARstar Omega plate reader (BMG Labtech) and normalised with respect to control reactions in the absence of compound ± Cp dimer (Full NCP = 1, degraded RNA = 0).