Journal: Nature protocols

Article Title: Rapid ordering of barcoded transposon insertion libraries of anaerobic bacteria

doi: 10.1038/s41596-021-00531-3

Figure Lengend Snippet: Three people work together to rapidly sort single cells. Station 1 is centered at the anaerobic chamber. The person working at Station 1 cycles between the anaerobic chamber and the cell sorter, bringing sets of fresh (unsorted) 96-deepwell plates to the cell sorter and taking back the set of sorted plates that have accumulated. Station 2 is next to the cell sorter. This person works to unseal fresh plates, pass them to Station 3, take sorted plates, and reseal them with a gas permeable membrane. Station 3 is the cell sorter. This person works to set up the equipment for the sort and run the single-cell sorting protocol. The arrows indicate movement of consumables between stations, and are color coded by the person doing the work.

Article Snippet: PlateOne 2-mL 96-deepwell polypropylene plates (USA Scientific, #1896–2000) Sterile 96-microwell V-bottom polystyrene plates (Grenier Bio-One, #651161) Sterile microwell plate lids (Grenier Bio-One, #656101) Standard profile polypropylene reservoirs (Mettler Toledo, # LR-R2-PB-5) Breathe-Easier gas permeable sealing membranes (Diversified Biotech, #BERM-2000) 5 mL polypropylene round bottom tubes (Falcon, #352063) Vinyl tape (3M, #471) Nunc Aluminum Seal Tape for 96-well plates (Thermo Scientific™, #232698) Alumaseal 384 films (Excel Scientific, #F-384–100) 1-mL LTS filter tips, for use with BenchSmart (Mettler Toledo, #30296782) 200-μL LTS filter tips, for use with BenchSmart (Mettler Toledo, #17010646) 200-μL filter tips, for use with multichannel pipette (Rainin #30389239) 8.5” x 8.75” OD PAKVF4 MylarFoil pouches, opening on the ZipSeal end (Impak Corporation, #085MFSOZE0875) 500-cc oxygen absorbing packets (Impak Corporation, #SF500CS1500) Microtube tough-tags for laser printers, 0.91×0.32 in (Diversified Biotech, #9186–1000) Microtube tough-tags for laser printers, 1.28×0.5 in (Diversified Biotech, #TTLW-2016) Glassware 2-L Pyrex bottles, 45-mm neck size (Pyrex, #1395–2L) 1-L Pyrex bottles, 45-mm neck size (Pyrex, #1395–1L) 250-mL Erlenmeyer flasks (Pyrex, #4980–250) SOFTWARE Software for random barcode transposon-site sequencing is available from https://bitbucket.org/berkeleylab/feba/src .

Techniques: FACS

Journal: Nature protocols

Article Title: Rapid ordering of barcoded transposon insertion libraries of anaerobic bacteria

doi: 10.1038/s41596-021-00531-3

Figure Lengend Snippet: Troubleshooting advice for common problems, with the associated step and possible explanation(s).

Article Snippet: PlateOne 2-mL 96-deepwell polypropylene plates (USA Scientific, #1896–2000) Sterile 96-microwell V-bottom polystyrene plates (Grenier Bio-One, #651161) Sterile microwell plate lids (Grenier Bio-One, #656101) Standard profile polypropylene reservoirs (Mettler Toledo, # LR-R2-PB-5) Breathe-Easier gas permeable sealing membranes (Diversified Biotech, #BERM-2000) 5 mL polypropylene round bottom tubes (Falcon, #352063) Vinyl tape (3M, #471) Nunc Aluminum Seal Tape for 96-well plates (Thermo Scientific™, #232698) Alumaseal 384 films (Excel Scientific, #F-384–100) 1-mL LTS filter tips, for use with BenchSmart (Mettler Toledo, #30296782) 200-μL LTS filter tips, for use with BenchSmart (Mettler Toledo, #17010646) 200-μL filter tips, for use with multichannel pipette (Rainin #30389239) 8.5” x 8.75” OD PAKVF4 MylarFoil pouches, opening on the ZipSeal end (Impak Corporation, #085MFSOZE0875) 500-cc oxygen absorbing packets (Impak Corporation, #SF500CS1500) Microtube tough-tags for laser printers, 0.91×0.32 in (Diversified Biotech, #9186–1000) Microtube tough-tags for laser printers, 1.28×0.5 in (Diversified Biotech, #TTLW-2016) Glassware 2-L Pyrex bottles, 45-mm neck size (Pyrex, #1395–2L) 1-L Pyrex bottles, 45-mm neck size (Pyrex, #1395–1L) 250-mL Erlenmeyer flasks (Pyrex, #4980–250) SOFTWARE Software for random barcode transposon-site sequencing is available from https://bitbucket.org/berkeleylab/feba/src .

Techniques: Microscopy, Incubation, Transferring, Sequencing