Finely ground leaves (100 g) were extracted with 90% methanol (MeOH) and stirred at room temperature. The resultant extracts were filtered through Whatman’s Grade 1 filter paper, and the collected filtrates were concentrated by using a rotary evaporator to afford a crude extract (12.4 g). This crude extract was suspended in distilled water (700 mL) and successively partitioned with the solvents ethyl acetate (EtOAc) and n-butanol (n-BuOH), thus yielding the EtOAc-soluble (8.3 g) and n-BuOH-soluble (1.5 g) fractions. The EtOAc-soluble fraction was further subjected to the Diaion HP-20 column in a gradient solvent system consisting of MeOH and H2O (100% H2O, 50% MeOH, 100% MeOH) to yield three subfractions W1 (2.0 g), W2 (2.5 g), and W3 (3.5 g). The W3 subfraction (100% MeOH) (3.5 g) was applied to Sephadex LH-20 column chromatography to give four successive fractions (W31‒W34). The W34 subfraction (2.7 g) was then fractionated by reversed-phase (RP)-silica column chromatography with 40% MeOH/H2O to afford five subfractions (W341‒W345). The W33 subfraction (352.5 mg) was separated using preparative RP HPLC with MeOH/H2O (gradient solvent system of 10–80% MeOH/H2O) to yield six subfractions (W331‒W336). Finally, the subfraction W334 (92.3 mg) was purified by the semi-preparative HPLC separation with 45% MeOH/H2O to isolated CDG (tR 12.8 min, 4.5 mg).
Grade 1 filter paper
Manufactured by Cytiva
Sourced in United Kingdom, Germany
Grade 1 filter paper is a laboratory filtration product used to separate solid particles from liquids. It is made of highly pure cellulose and provides reliable and consistent filtration performance.
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Lab products found in correlation
Bovine liver powder, by MP Biomedicals (2 mentions)
Artificial blood feeding system, by Hemotek (2 mentions)
Fei vitrobot mk 4, by Thermo Fisher Scientific (1 mentions)
Gf c glass microfiber filter paper, by Cytiva (1 mentions)
3mm chr chromatography paper, by Cytiva (1 mentions)
Rotary evaporator, by Heidolph (1 mentions)
Methanol, by Merck Group (1 mentions)
Hexane, by Merck Group (1 mentions)
Human blood, by Hemotek (1 mentions)
Rotavapor r 200, by Büchi (1 mentions)
Rotavapor r 210, by Büchi (1 mentions)
Snap ultra c18 cartridge, by Biotage (1 mentions)
Isolera one, by Biotage (1 mentions)
Eagle 4k ccd camera, by Thermo Fisher Scientific (1 mentions)
T12 electron microscope, by Thermo Fisher Scientific (1 mentions)
Drp 110, by Metrohm (1 mentions)
Alpha q water system, by Merck Group (1 mentions)
Drp dsc, by Metrohm (1 mentions)
Gp50 gradient pump, by Thermo Fisher Scientific (1 mentions)
Carbopac pa1 column, by Thermo Fisher Scientific (1 mentions)
49 protocols using grade 1 filter paper
1
Isolation and Purification of CDG from Plant Extract
2
Green Synthesis of Silver Nanoparticles
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The AgNPs were synthesized by a simple one-pot green route method as shown in figure 1 b. The fresh leaves of Pistia stratiotes (30 g) were cut into small pieces and cleaned by DI water. Then, they were heated in 500 ml of DI water at 95°C for 30 min and cooled down to room temperature. The reducing solution was filtered by Whatman® Grade 1 filter paper. Next, 0.04 g of silver nitrate (AgNO3) in 20 ml DI water was filled into the reducing P. stratiotes solution (100 ml). To reduce Ag ion into AgNPs, the mixture solutions were put under a blue-light illumination (8 W lamp) for 1 h in a dark chamber at room temperature. Successful formation of AgNPs can be observed by colour change from yellow to brown with the naked eye.
3
Methanolic Extraction of Plant Compounds
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Two grams of seeds from each sample were extracted on a magnetic shaker for 3 h using 25 mL of methanol (80%) as the solvent. The resulting solution was filtered through a Whatman Grade 1 filter paper, and after it was centrifuged at 3000 rpm for 20 min, the supernatant was stored at -80 °C until the experiment day.
4
Phenolic Compound Extraction and Analysis
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Three sub-samples of 25 g each were taken from the raw and cooked samples; after homogenization, they were extracted (twice for 1 h) under reflux in a hot water bath with boiling methanol (1:10 w/v). The combined methanolic extracts were concentrated under reduced pressure by a rotary evaporator. The obtained residue was dissolved in 50% (v/v) methanol/water; the resulting solution was filtered through a Whatman Grade 1 filter paper and used for: (i) the evaluation of total phenolic concentration; (ii) the HPLC analysis of phenolic compounds; and (iii) the antioxidant activity assay. All details of the extraction procedure were as already reported by Sergio et al. [14 (link)].
5
Cryo-EM analysis of ATM-p53 complex
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Samples (20 μl) were prepared in PCR tubes with 2 μM ATM Q2971A (with N-terminal FLAG-FKBP tag), 0.5 mM p53 peptide (residues 11 to 22), 1 mM H2O2, 1 mM AMP-PNP, and 5 mM MgCl2 using a dilution buffer of 50 mM Hepes (pH 7.5), 100 mM NaCl, and 10% glycerol. Samples were first incubated with all components except H2O2. Tubes were kept on ice for around 30 min while setting up an FEI Vitrobot Mk IV and glow discharging grids. The Vitrobot chamber was set to 30°C and 100% humidity. UltrAuFoil R1.2/1.3 Au 300 mesh grids were glow-discharged using a sputter coater discharger (Edwards, S150B) for 1 min and 15 s at 0.8 kV, 30 mA, and 10−1 torr. Sample tubes were moved to an RT holder and incubated at RT for 1 min; then, 2 μl of a 10 mM H2O2 stock was added to the tubes. Sample tubes were further incubated at RT between 7.5 and 15 min, and 4-μl droplets were pipetted onto grids. Grids were blotted using Whatman grade 1 filter paper with 0-s incubation time, 2-s blot time, and −7 blot force and vitrified in liquid ethane using an FEI Vitrobot Mk IV.
6
RNAi Experiments in Paramecium
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In all RNAi experiments, P. bursaria 186b (CCAP 1660/18) strain was used. For experiments requiring a nonphotoendosymbiotic Paramecium system for comparison, P. tetraurelia nd7 strain was used. For eDicer analysis, P. bursaria 186b and Yad1g1N strains were used (42 (link), 60 (link)).
Paramecium cells were cultured in New Cereal Leaf Prescott Liquid media (NCL). NCL media was prepared by adding 4.3 mg ⋅ L−1 CaCl2.2H2O, 1.6 mg ⋅ L−1 KCl, 5.1 mg ⋅ L−1 K2HPO4, and 2.8 mg ⋅ L−1 MgSO4.7H2O to deionized water. Then, 1 g ⋅ L−1 wheat bran was added, and the solution was boiled for 5 min. Once cooled, media was filtered once through Whatman Grade 1 filter paper and then through Whatman GF/C glass microfiber filter paper. Filtered NCL media was autoclaved at 121 °C for 30 min to sterilize prior to use.
NCL medium was bacterized with Klebsiella pneumoniae SMC and supplemented with 0.8 mg ⋅ L−1 β-sitosterol prior to propagation. Paramecium cells were subcultured 1:9 into fresh bacterized NCL media once per month for P. bursaria 186b and once every 2 wk for P. tetraurelia nd7. Paramecium cultures were maintained at 18 °C with a light–dark cycle of 12:12 h.
Paramecium cells were cultured in New Cereal Leaf Prescott Liquid media (NCL). NCL media was prepared by adding 4.3 mg ⋅ L−1 CaCl2.2H2O, 1.6 mg ⋅ L−1 KCl, 5.1 mg ⋅ L−1 K2HPO4, and 2.8 mg ⋅ L−1 MgSO4.7H2O to deionized water. Then, 1 g ⋅ L−1 wheat bran was added, and the solution was boiled for 5 min. Once cooled, media was filtered once through Whatman Grade 1 filter paper and then through Whatman GF/C glass microfiber filter paper. Filtered NCL media was autoclaved at 121 °C for 30 min to sterilize prior to use.
NCL medium was bacterized with Klebsiella pneumoniae SMC and supplemented with 0.8 mg ⋅ L−1 β-sitosterol prior to propagation. Paramecium cells were subcultured 1:9 into fresh bacterized NCL media once per month for P. bursaria 186b and once every 2 wk for P. tetraurelia nd7. Paramecium cultures were maintained at 18 °C with a light–dark cycle of 12:12 h.
7
Extraction of P. dodecandra Root Powder
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Three hundred grams of the coarse powder of the roots of P. dodecandra were macerated for 3 days using 80% methanol as a solvent. Then, the fluid extract was filtered using muslin cloth and Whatman® grade 1 filter paper and the marc was re-extracted for the second and third times by adding another fresh 80% methanol. The extract was concentrated in a rotary evaporator (Buchi, Rotavapor R-210/215, Switzerland), freeze-dried in a lyophilizer (OPERON, OPR-FDU-5012, Korea), and stored in a chest freezer.
8
Quantifying Phenolics and Antioxidant Activity
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Juice samples (10 g) were extracted twice with 20‐ml methanol on a shaker for 3 hr. The extracts from each sample were pooled, filtered using Whatman grade 1 filter paper, and further used for quantification of total phenolics and radical scavenging activity using a DPPH assay. Volume of the extracts was measured for calculating dilution factor.
9
Point-of-Care DNA Detection Device
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Each device contains four panels, namely: an absorbent pad (Panel A, 1.5 cm × 1.5 cm) for sample purification and washing; Whatman 3MM CHR chromatography paper (Panel B, with a diameter of 4 mm) for cell lysis; Whatman Grade 1# filter paper (Panel C, with a diameter of 4.5 mm) for DNAzyme recognition, and Nitrocellulose membrane HF180 (Panel D, with a diameter of 4.5 mm) for RCA. For dry reagent storage, 5 µL of cell Lysis Buffer was pipetted onto the Panel B. The Panel was dried in a vacuum oven at 50 °C for about 1 h. For Panel C, 5 μL of 3D EC1 was printed using a Biodot XYZ3060 automated dispensing unit (Somerville, MA, USA). After immersion into the blocking buffer (25 mM PBS containing 1% BSA, 0.02% Tween-20, pH 7.5) for 10 min, the obtained bioactive paper was dried at RT. 5 µL of F-RS28 (100 nM) was then mixed with 5 µL of 10% (w/v) pullulan, and then printed onto Panel C using the dispensing unit. For Panel D, 1 µL of 1 µM CDT2, 1 µL of 10 mM dNTPs, 1 µL of 100 µM hemin, 4 U of PNK and 5 U of ϕ29DP were first mixed with 20 µL of 10% (w/v) pullulan, and then printed. The bioactive paper was dried and stored at RT in a desiccant container. In order to assemble these four panels, a single sided adhesive film was used to enable bidirectional folding.
10
Synthesis of Silver Nanoparticles from Acer oblongifolium Leaves
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Acer oblongifolium fresh leaves were cleaned by washing under tap water and then rinsed with deionized water. The leaves were then shadow dried for 2–3 days and ground into powder using an electric blender. Leaf powder of 4 g was suspended in distilled water (200 mL) and kept in a water bath at a temperature of 70–80 °C for 30 min in a beaker. The remaining extracts were filtered in a conical flask using Whatman Grade 1 Filter Paper, cooled down, and refrigerated at 4 °C for further use in the synthesis of AgNPs according to the method described by Krithiga et al. and Pirtarighat et al. [26 (link),27 (link)].
AgNO3 (25 mM, 1 M) solution was made by mixing AgNO3 flakes in deionized water. This solution was used for synthesizing AgNPs with the aim of testing their antibacterial activity.
AgNO3 (25 mM, 1 M) solution was made by mixing AgNO3 flakes in deionized water. This solution was used for synthesizing AgNPs with the aim of testing their antibacterial activity.
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