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. 2020 Jan 16;16(1):e9111. doi: 10.15252/msb.20199111

Figure 1. A schematic overview of automated single‐pot solid‐phase‐enhanced sample preparation (autoSP3) workflows.

Figure 1

  1. The overview shows the different steps of the autoSP3 protocol from protein input to enzymatic digestion. The setup of the Bravo deck is shown for the core clean‐up protocol.
  2. The overview shows the steps and Bravo deck setup of the autoSP3 peptide acidification and recovery protocol. The protocol ends with MS injection‐ready peptide samples.
  3. A schematic overview of all available autoSP3 protocol versions. The autoSP3 procedure is provided with three options for reduction and alkylation and with post‐digestion peptide recovery as described in the Appendix Protocols (A to D). Protocol A: one‐step reduction and alkylation using a TCEP/CAA mixture for 5 min at 95°C, followed by autoSP3. Protocol B: two‐step reduction and alkylation using DTT and CAA consecutively with 30 min of incubation at 60 and 23°C, respectively, followed by autoSP3. Protocol C: the core autoSP3 protocol omitting reduction and alkylation such that the user can flexibly pre‐treat manually prepared samples. Protocol D: post‐digestion acidification and recovery, delivering MS injection‐ready peptides to a new sample plate.