Skip to main content
. Author manuscript; available in PMC: 2022 Apr 1.
Published in final edited form as: Nat Protoc. 2021 Aug 11;16(10):4676–4691. doi: 10.1038/s41596-021-00592-4

Fig. 1 |. MetaRibo-Seq protocol workflow.

Fig. 1 |

A biopsy punch with a plunger is used to prepare 300 mg aliquots of stool into two screw-top tubes (150 mg per tube) with lysis buffer (recipe 1). Bacterial cells are lysed using a bead beater and centrifuged to pellet fecal debris, and an ethanol precipitation is performed on the supernatant. Precipitate is resuspended in MNase buffer (recipe 2) and digested with MNase. Monosomes are recovered using size-exclusion columns, purified, and subjected to rRNA depletion. MetaRibo-Seq libraries are created from these footprints and sequenced.