Improve Research Reproducibility A Bio-protocol resource
Concise Method
tooltip

Growth experiments for determining iron utilization

AM Avishek Mitra
YK Ying-Hui Ko
GC Gino Cingolani
MN Michael Niederweis
request Request a Protocol
ask Ask a question
Favorite

Strains were first grown in 7H9 medium, then washed in sterile PBS (137 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 1.8 mM KH2PO4) with 0.02% Tyloxapol and iron-depleted for 3–4 generations in iron-free 7H9 medium. Strains were then inoculated into HdB minimal medium60 containing either 10 µM hemin or 2.5 µM human hemoglobin or 10 µM ammonium ferric citrate as the sole iron source. BSA was added to a final concentration of 0.5% w/v into HdB minimal medium for albumin growth experiments. To prevent growth from iron traces 20 µM of the iron chelator 2,2-dipyridyl (DIP) was added to the medium containing heme or human hemoglobin.

Copyright and License information: The Author(s) ©2019
Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

Do you have any questions about this protocol?

Post your question to gather feedback from the community. We will also invite the authors of this article to respond.

post Post a Question
0 Q&A