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Berberine chloride preparation.

JW Judy J. Wan
RB Rebecca S. Brown
MK Margaret Kielian
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Berberine chloride (BBC) was purchased from Sigma-Aldrich (catalog no. B3251) and stored in the dark under desiccating conditions. Fresh stock solutions of BBC were prepared for each experiment. For cell culture experiments, 50 mM BBC stock was prepared in sterile, tissue culture-grade 100% dimethyl sulfoxide (DMSO), incubated in the dark for 30 min at 37°C with intermittent vortexing, and then sonicated at room temperature for two bursts of 2 min each in a bath sonicator (Avanti). For in vitro experiments, 2.5 mM BBC was prepared in molecular grade water to avoid potential denaturing effects of DMSO on RNA. The stock was incubated at 37°C for 1 h with intermittent vortexing, filtered through a 0.22-μm syringe filter, and buffered to a final concentration of 2.5 mM BBC in 25 mM K-HEPES (pH 7.4) and 100 mM KCl immediately before addition to in vitro reaction mixtures.

Copyright and License information:  ©2020 Wan et al.
This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.

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