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MB Sample ID: SA220010
| Local Sample ID: | YL58.3 |
| Subject ID: | SU002375 |
| Subject Type: | Bacteria |
| Subject Species: | Several mouse commensals |
| Age Or Age Range: | 8-16 hours |
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Collection:
| Collection ID: | CO002368 |
| Collection Summary: | Bacteria were grown in 15mL culture tubes in 3mL of modified brain heart infusion broth [37g/L BHI powder, 0.025% Cystiene-HCl.H2O, 0.025% Na2S.9H2O, 1ug/mL Hemin, 0.5ug/mL menadione, 0.025% mucin]. Media was pre-reduced in an anaerobic chamber (Whitley A95 Workstation; 90% N2, 5% H2 & 5% CO2) for 48 hours then bacterial cultures were started from glycerol stocks. 200µL of glycerol stock was added to each culture tube and cultures were incubated for 6-36 hours until early plateau phase. After incubation, cultures were removed from the anaerobic chamber, and 50uL was collected to extract DNA for 16S sequencing to confirm accurate identification of bacteria and absence of contamination. The remaining culture was centrifuged (max. speed, 15 minutes, 4°C) and the supernatant was collected and combined 1:1 with 100% methanol. Samples were then incubated at - 20°C for 1 hour. Samples were centrifuged (max. speed, 15 minutes, 4°C), supernatant was recovered and combined 1:4 with 50% methanol to obtain a final dilution of D20. |
| Sample Type: | Bacterial cells |
| Storage Conditions: | Described in summary |
