Clostridioides difficile, an obligate anaerobic bacterium, causes infections leading to prolonged diarrhea. The bacterium produces dormant spores that can withstand an aerobic environment, resulting in easy environmental transfer. Here, we present a convenient sporulation and purification protocol that can be practiced in any lab setting using a portable anaerobic glove bag. This protocol also optimizes existing cell growth methods and presents a detailed trouble shooting guide. This protocol is a modification of those previously reported by Edwards and McBride (2016) and Shen et al. (2016).
Bibliographical noteFunding Information:
This work was supported, in part, by the Office of the Assistant Secretary of Defense for Health Affairs , under Investigator Initiated Research Awards W81XWH-17-1-0635 (P.I.D.) and W81XWH-17-1-0636 (A.K.) and by grant 5U01HL127479-03 from the NIH , under the MIN-REACH Research and Evaluation Hub. We would like to thank Alexa Weingarden and Raymond Erickson for helpful discussions and John Ferguson for help with images.
© 2020 The Author(s)
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