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Samples of this mixture were collected using a precipitation funnel where components of the mixed plumes were discharged from the atmosphere with the rainfall.
To analyze the formation of stable complexes between KaiB and KaiC, KaiB (50 ng/µl) and KaiC (200 ng/µl) were mixed and incubated at 30°C in a volume of 100 µl RB. 16 µl aliquots of each mixture were collected at each time point and mixed with 5X non-denaturing buffer (0.312 M Tris-HCl, pH 6.8, 0.05% CBB, 50% glycerol) and frozen immediately at −80°C.
Nanofractions of the standard mixture were collected in two relatively low (24 and 12 s/well) and two high (6 and 3 s/well) resolution.
Briefly, donor and recipient strains were mixed together in a 10 1 v/v ratio and cells in 50 μl volumes of each mixture were collected by vacuum filtration onto a sterile 0.22-μm filter disc.
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After 5, 10, 20, and 30 min incubation, 5 mL of the mixture was collected and centrifuged, followed by collection of the pellet (5, 10, 20, and 30 min samples).
Then, the effluent gas mixture was collected for chromatographic partitioning analysis.
At predetermined times, 5 ml of reaction mixture was collected and centrifuged (4000 rpm, 15 min) in a centrifuge.
Approximately 20 μL of each mixture was collected after 2, 3 and 9 days of incubation and analysed by SDS-PAGE on a NuPAGE 4% to 12% Bis-Tris gel (Invitrogen; Carlsbad, CA, USA) in MES buffer (200 V constant, approximately 30 min).
The bead/conidia mixture was collected into a tube and suspended in 1 mL sterile phosphate buffered saline (PBS).
The emulsified mixture was collected into an airtight vial (containing 500 µL buffer to dilute and thus prevent protein synthesis from taking place in any cell-free mixture that avoids emulsification) and returned to the glove box immediately upon completion.
After the purified PrC was incubated with the nematode cuticle (1 mg/ml) for 20 min, the reaction was terminated by heat at 100°C for 5 min. The reaction mixture was collected and applied to a centrifugal iCONTM concentrator (MWCO 3 kD, Pierce, Rockford, IL).
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