Bacterial cell pellets were frozen in liquid nitrogen and lyophilized to dryness. One milliliter of HPLC-grade methanol was added to the dried material, and the mixture was sonicated for 3 minutes (on/off pulse cycles of 2 seconds on, 2 seconds off, at power 100%) using a Qsonica Ultrasonic Processor (Model Q700) with a water bath cup horn adapter (Model 431C2), with water bath flow to maintain room temperature. Samples were then moved to an end-over-end rotator, and extractions proceeded for 12 hours. Samples were then centrifuged at 18,000 × g for 30 minutes at 4°C. The supernatant was transferred to a fresh centrifuge tube, and the solvent was dried with a Thermo Scientific Savant SpeedVac SPD130DLX. The dried material was resuspended in 200 µL HPLC-grade methanol, briefly sonicated, and centrifuged as before. The concentrated extract was transferred to an HPLC vial with a 300-µL glass insert and stored at 4°C until further analysis.
Savant speedvac spd130dlx
Manufactured by Thermo Fisher Scientific
The Savant SpeedVac SPD130DLX is a centrifugal vacuum concentrator designed for laboratory applications. It is capable of concentrating and drying samples by evaporating solvents at reduced pressure and controlled temperature.
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Lab products found in correlation
2 protocols using savant speedvac spd130dlx
1
Bacterial Metabolite Extraction Protocol
2
Metabolite Extraction for LC-MS Analysis
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Samples were frozen over liquid nitrogen and lyophilized to dryness. 1 mL of HPLC grade methanol was added to the dried material and the mixture was sonicated for 3 min (on/off pulse cycles of 2 second on, 2 seconds off, at power 100%) using a Qsonica Ultrasonic Processor (Model Q700) with a water bath cup horn adaptor (Model 431C2), with water bath flow to maintain approximately room temperature. Samples were then moved to an end-over-end rotator and extractions proceeded for 12 hours. Samples were then centrifuged at 18000 G for 30 minutes at 4 °C. The supernatant was transferred to a fresh centrifuge tube and solvent was dried with a Thermo Scientific Savant SpeedVac SPD130DLX. The dried material was resuspended in 200 μL HPLC-grade methanol, briefly sonicated, and centrifuged as before. The concentrated extract was transferred to HPLC vial with a 300 μL glass insert and stored at 4 °C until further analysis.
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