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MB Sample ID: SA187687

Local Sample ID:	c14
Subject ID:	SU002085
Subject Type:	Other abiotic sample

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Sample Preparation:

Sampleprep ID:	SP002091
Sampleprep Summary:	For every sample, four aliquots with 0.75 mL of sludge were prepared in 2-mL Eppendorf tubes. Then, 1 mL of phosphate buffer saline was added into each tube and the sludge volumes were washed by centrifuging them at 15500 rcf for 3 min at 4 °C. This process was repeated once. The samples were freeze-dried overnight and, for every sample, 20 mg of dried sludge were transferred in new Eppendorf tubes. Then, the dried sludge powders were resuspended in 1 mL of a cold ddH2O: MeOH mixture (1:1) containing 2.5 mM PIPES. The samples were sonicated for 6 min, and submerged in liquid nitrogen. After one minute, the samples were thawed in ice. The cold-shock step was repeated twice. Afterwards, the samples were agitated for 15 min at 420 rpm in an orbital shaker. Following this, the samples were centrifuged at 15500 rcf for 3 min at 4 °C and the supernatants were collected. The supernatants were immediately filtered with 0.45 μM Nylon filters and freeze-dried overnight. The dried extracts were left at -80°C until the HPLC-MS analyses.

Sampleprep ID:

SP002091

Sampleprep Summary:

For every sample, four aliquots with 0.75 mL of sludge were prepared in 2-mL Eppendorf tubes. Then, 1 mL of phosphate buffer saline was added into each tube and the sludge volumes were washed by centrifuging them at 15500 rcf for 3 min at 4 °C. This process was repeated once. The samples were freeze-dried overnight and, for every sample, 20 mg of dried sludge were transferred in new Eppendorf tubes. Then, the dried sludge powders were resuspended in 1 mL of a cold ddH2O: MeOH mixture (1:1) containing 2.5 mM PIPES. The samples were sonicated for 6 min, and submerged in liquid nitrogen. After one minute, the samples were thawed in ice. The cold-shock step was repeated twice. Afterwards, the samples were agitated for 15 min at 420 rpm in an orbital shaker. Following this, the samples were centrifuged at 15500 rcf for 3 min at 4 °C and the supernatants were collected. The supernatants were immediately filtered with 0.45 μM Nylon filters and freeze-dried overnight. The dried extracts were left at -80°C until the HPLC-MS analyses.