PEGylated cypate in methanol was dried under rotary evaporation, and the resulting thin film was hydrated with 1 ml of PBS (pH 7.4) and vortexed. After sonication for 10 min, large aggregates were removed by filtration through 0.2-μm polycarbonate membrane filters (Millipore Corp., Billerica, MA), and SP3NPs were obtained and stored at 4 oC until use. The size and morphology of SP3NPs were examined by TEM using a JEM1010 system (Jeol Ltd, Tokyo, Japan). The hydrodynamic diameters and zeta potentials of SP3NPs were measured using a Nanosizer ZS90 instrument (Malvern Instruments Ltd, Malvern, UK). The colloidal stability of SP3NPs in PBS and 50% FBS was tested by monitoring changes in size over 1 wk. The UV-Vis spectra of SP3NPs were recorded using a UV-Vis spectrophotometer (NEOSYS2000; Scinco, Twin Lakes, WI).
0.2 μm polycarbonate membrane filters
Manufactured by Merck Group
Sourced in Morocco
The 0.2-μm polycarbonate membrane filters are designed for the filtration of a variety of liquids. These filters have a pore size of 0.2 micrometers, which allows for the removal of particulates, microorganisms, and other contaminants from the sample.
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2 protocols using 0.2 μm polycarbonate membrane filters
1
Characterization of Stabilized Fluorescent Nanoparticles
2
Microbial Community Analysis via 16S rDNA Sequencing
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Subsamples of 150 mL were filtered onto 0.2 μm polycarbonate membrane filters (47 mm diameter, Millipore), transferred to 2 mL sterilized microcentrifuge tubes and stored at -20°C for subsequent molecular analysis. Molecular analysis of microbial communities was performed according to previous studies [33] . In brief, microbial DNA was extracted from frozen filters using the E.Z.N.A.® Water DNA Kit (Omega, USA) according to manufacturer's protocols. Amplicons of V3-V4 regions of the 16S rDNA gene were sequenced on the Illumina HiSeq platform at Shanghai Xiangyin Biotechnology Co., using the universal primer set SD-Bact-0341-b-S-17/SD-Bact-0785-a-A-21 (5 ′ -CCTACGGGNGGCWGCAG-3 ′ / 5 ′ -GACTACHVGGGTATCTAATCC-3 ′ ) [34] . Denoised sequences were aligned and clustered at 97% sequence identity into operational taxonomic units (OTUs), and these OTUs were assigned taxonomic identities using the Ribosomal Database Project (RDP) classifier [35] . Alpha-diversity measures (observed OTUs, Good's coverage, abundance-based coverage estimator (ACE), Chao1 richness estimator, Shannon index and Simpson index) were calculated based on OTU data. All sequence data from this study were submitted to the National Center for Biotechnology Information (NCBI) Sequence Read Archive (SRA) under accession number SRP148474.
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