Whatman filter paper #1 was purchased from GE Healthcare Life Sciences. Wax paper squares (6″) were purchased from NORPRO. Gold (III) chloride hydrate, sodium citrate tribasic, poly (ethylene glycol) 2-mercaptoethyl ether acetic acid (thiol-PEG-acid) 2100, N-hydroxysulfosuccinimide sodium salt (sulfo-NHS), N-(3-Dimethylaminopropyl)-N′- ethylcarbodiimide hydrochloride (EDAC; EDC), poly (sodium 4-styrenesulfonate, average Mw ∼70.000) 30 % solution, polyethylene glycol 3000 (biotin-PEG), Tween-20, 2-ethanesulfonic acid (MES), and anti-mouse IgG (Fc Specific) (SAB3701021) were obtained from Sigma Aldrich. Mouse monoclonal antibody Anti-SARS-CoV-2 N-protein (35–579) was purchased from ProScience. Recombinant Nucleocapsid protein from SARS-CoV-2 (230−01104) was purchased from Raybiotech. Bovine serum albumin (BSA, protease free) was purchased from VWR chemicals. Aerosol bottles were purchased from Lurrose. Surgical facemasks for experiments in Fig. 3 were purchased from Medline (NONE27377). PBS refers to phosphate buffered saline pH 7.4. PBST refers to PBS supplemented with 0.1 % Tween-20. RT refers to room temperature. PBS-BSA refers to PBS supplemented with 0.5 % BSA.
Whatman 1 filter paper
Manufactured by GE Healthcare
Sourced in United States
Whatman #1 filter paper is a high-quality cellulose-based filtration product commonly used in various laboratory applications. It is designed to effectively separate solid particles from liquids, enabling efficient filtration processes.
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Lab products found in correlation
Whatman, by Cytiva (8 mentions)
Sodium citrate tribasic, by Merck Group (1 mentions)
2 ethanesulfonic acid mes, by Merck Group (1 mentions)
Gold 3 chloride hydrate, by Merck Group (1 mentions)
Yeast extract, by Merck Group (1 mentions)
Potassium chloride (kcl), by Merck Group (1 mentions)
Milli q water purification system, by Merck Group (1 mentions)
Kh2po4, by Merck Group (1 mentions)
Ampicillin, by Merck Group (1 mentions)
Na2hpo4, by Merck Group (1 mentions)
Bacitracin, by Merck Group (1 mentions)
Resazurin sodium salt, by Merck Group (1 mentions)
Gentamicin, by Merck Group (1 mentions)
Tryptone, by Merck Group (1 mentions)
Erythromycin, by Merck Group (1 mentions)
Cellulose filters paper mn 615 no 1, by Macherey-Nagel (1 mentions)
T4 dna ligase, by Takara Bio (1 mentions)
Bca protein assay kit, by Beyotime (1 mentions)
29 dna polymerase, by Thermo Fisher Scientific (1 mentions)
Whatman 3mm chr chromatography paper, by GE Healthcare (1 mentions)
8 protocols using whatman 1 filter paper
1
Development of SARS-CoV-2 Antigen Assay
2
Antibiotic Sensitivity Testing Procedure
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Tetracycline was obtained from Boehringer Mannheim GmbH/Roche Diagnostics GmbH (Germany). Gentamicin, erythromycin, ampicillin, bacitracin, resazurin sodium salt and KCl were supplied by Sigma (St. Louis, MA, USA). Cellulose filters (Paper) (MN 615/No 1) were obtained from Macherey-Nagel (Düren, Germany). Acetonitrile, HPLC-grade far UV cut-off was supplied by Romil Ltd (Cambridge, UK). Merck (Darmstadt, Germany) supplied agar, yeast extract, tryptone, Na2HPO4, KH2PO4 and Merck (Wadeville, SA) supplied sodium chloride. Petri dishes were provided by Greiner bio-one (Frickenhausen, Germany). The 96-well polystyrene plates were acquired from Corning (Kennebunk, ME, USA) and Whatman Filter paper 1 from GE Healthcare Life Sciences supplied by Sigma-Aldrich (Darmstadt, Germany). Analytical grade water (milliQ water) was obtained by filtering water in a reverse osmosis plant through Millipore Milli-Q® water purification system (Milford, USA).
3
Oligonucleotide Purification and Characterization
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All DNA oligonucleotides were obtained from Takara Biotechnology Co. (Dalian, China) and purified by standard 10% denaturing (8 M urea) polyacrylamide gel electrophoresis (dPAGE) or high-performance liquid chromatography (HPLC). T4 DNA ligase, dNTP, and T4 polynucleotide kinase (PNK) were purchased from Takara Biotechnology Co. (Dalian, China). ϕ29 DNA polymerase was acquired from Thermo Scientific (Waltham, MA, USA). Pullulan was obtained from Sangon Biotech (Shanghai, China). BCA Protein Assay Kit was acquired from Beyotime Biotechnology (Shanghai, China). Triton X-100 cell lysis buffer SS0890 was obtained from NOVON (Beijing, China). Nitrocellulose paper (HF180), Whatman filter paper 1# and Whatman 3MM CHR chromatography paper were purchased from GE Healthcare (Chicago, IL, USA). All other chemicals were purchased from Sangon Biotech (Shanghai, China) and used without further purification. Water was purified with a Greate Fun SUMMER-S-10 water purification system.
The sequences of the DNA molecules were as follows:
CDT1 (anti-EC1): GATATCATAT CACACAACTG TAAAGAAATC CATCCCCACA CAGTGTAGTG TTCCGGTCGC AGGTCTCGAC AACGCACATC (5′→3′)
TP1: ATATGATATC GATGTGCGTT (5′→3′)
DT1: ACGCACATCG ATATCATATC AC (5′→3′)
CDT2: TAGCTAGGAA GAGTCCCAAC CCGCCCTACC CAAAATGTCT CGGAT (5′→3′)
TP2: TTCCTAGCTA ATCCGAGACA (5′→3′)
F-RS28: ACTCTTCCTA GCFrAQGGTTC GATCAAGA-invert dT (5′→3′)
The sequences of the DNA molecules were as follows:
CDT1 (anti-EC1): GATATCATAT CACACAACTG TAAAGAAATC CATCCCCACA CAGTGTAGTG TTCCGGTCGC AGGTCTCGAC AACGCACATC (5′→3′)
TP1: ATATGATATC GATGTGCGTT (5′→3′)
DT1: ACGCACATCG ATATCATATC AC (5′→3′)
CDT2: TAGCTAGGAA GAGTCCCAAC CCGCCCTACC CAAAATGTCT CGGAT (5′→3′)
TP2: TTCCTAGCTA ATCCGAGACA (5′→3′)
F-RS28: ACTCTTCCTA GCFrAQGGTTC GATCAAGA-invert dT (5′→3′)
4
Biomass Pretreatment and Extractives Analysis
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Pretreated but unwashed biomass (equivalent to 1 g glucan according to Table 2 ) was washed sequentially with 20 ml of water, 20 ml ethanol, and 20 ml acetone on a Buchner funnel through Whatman #1 filter paper (GE Life Sciences, Piscataway, NJ, United States). The resulting extracted material solutions were dried at 50°C overnight and redissolved in 10 ml water. Glc content in the extractives was assayed directly as described above. The extractives were then digested to completion with C/HTec2 and re-measured for free Glc. The difference before and after enzymatic digestion was taken as Glc due to soluble oligosaccharides, with a small contribution from the C/HTec2 itself, which was subtracted.
See Materials and Methods for the GenBank or JGI accession numbers.
Enzyme inhibition by the extractives was tested in a standardized assay containing CS at a concentration of 2 mg glucan/ml and a C/HTec2 loading of 1 mg/g glucan, as described below. Extractives derived from the equivalent of 1 mg glucan from unwashed biomass were tested per mg of glucan from corn stover.
See Materials and Methods for the GenBank or JGI accession numbers.
Enzyme inhibition by the extractives was tested in a standardized assay containing CS at a concentration of 2 mg glucan/ml and a C/HTec2 loading of 1 mg/g glucan, as described below. Extractives derived from the equivalent of 1 mg glucan from unwashed biomass were tested per mg of glucan from corn stover.
5
Mushroom Bioactive Compound Extraction
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Mushroom fruiting bodies were cleaned using a brush to remove dirt, cut into small pieces, air-dried at 25°C—30°C for 5–7 days and pulverized using a blender. Ten grams of each powdered mushroom was mixed with 100 mL 95% ethanol, tumbled at room temperature for 3 days, centrifuged at 1000 X g for 10 min and the resulting supernatant was filtered using Whatman #1 filter paper (GE Healthcare Life Sciences, Piscataway, NJ, USA). Each filtrate was vacuum-dried using Eppendorf Vacufuge Plus Concentrator System (Eppendorf, Enfield, CT, USA). The dried extracts were tared to constant weight and dissolved in dimethyl sulfoxide (DMSO) to prepare 50,000 ppm (parts per million) stock solutions.
6
Seed Germination and Seedling Growth Protocol
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Twenty large seeds or 0.5 g of small seeds of each accession were put in sterile 15- or 2-ml tubes and soaked for 6 h in double-distilled sterile water (Figure 1C ). Half of these were then transferred to a sterile Petri dish containing a sterile Whatman #1 filter paper (GE HealthCare: United States) and irrigated with 3 ml of sterile water, while the other half received 3 ml of sterile water mixed with 1 g of field soil. These were incubated at 32°C in the dark for several days until germination.
From each dish, two seedlings were transplanted to corresponding jars as they germinated. Autoclaved glass jars were 13-cm tall, 7-cm wide in diameter, and filled with 100 ml of sterile sand (then autoclaved again) or with 100 ml of 1:1 soil/sterile sand, then watered once with 10 ml of sterile distilled water and sealed with a plastic lid. Jars were incubated (and never opened) in a single Panasonic MLR-352H Plant Growth Chamber set at 28°C for 12 h with 5 lm of fluorescent light and for 12 h of darkness at 22°C. Plants were grown between 2 weeks and 2 months, until they were of a significant size or until they hit the lid of the jar. Before harvesting, jar lids were removed inside a laminar flow hood, and shoots were allowed to dry off for 24 h (Figures 1D,E ).
From each dish, two seedlings were transplanted to corresponding jars as they germinated. Autoclaved glass jars were 13-cm tall, 7-cm wide in diameter, and filled with 100 ml of sterile sand (then autoclaved again) or with 100 ml of 1:1 soil/sterile sand, then watered once with 10 ml of sterile distilled water and sealed with a plastic lid. Jars were incubated (and never opened) in a single Panasonic MLR-352H Plant Growth Chamber set at 28°C for 12 h with 5 lm of fluorescent light and for 12 h of darkness at 22°C. Plants were grown between 2 weeks and 2 months, until they were of a significant size or until they hit the lid of the jar. Before harvesting, jar lids were removed inside a laminar flow hood, and shoots were allowed to dry off for 24 h (
7
Metabolite Extraction from Fungal Cultures
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We prepared metabolite extracts used for bioassays from whole plate cultures of 11-1-2 by cutting the agar into pieces and transferring the pieces into a clean flask. Ethyl acetate (20 mL) was added to each flask, and the contents were mixed and left to incubate at room temperature overnight. The extracts were then filtered using Whatman #1 filter paper (GE Healthcare Life Sciences) and transferred into clean flasks. The remaining agar pieces were rinsed with 10 mL of fresh ethyl acetate, which was subsequently filtered and combined with the corresponding extract. The solvent was evaporated overnight, and the dried extracts were resuspended in 1 mL of 100% vol/vol LC-grade methanol and were stored at −20°C.
For the metabolomic analysis of 11-1-2, organic extractions were carried out as described above except that the solvent was removed by rotary evaporation, and the dried extracts were redissolved in 1 mL of 100% vol/vol LC-MS-grade methanol. A 500-μL aliquot of each extract was then transferred into a 96-well plate and used for LC-MS/MS analysis.
For the metabolomic analysis of 11-1-2, organic extractions were carried out as described above except that the solvent was removed by rotary evaporation, and the dried extracts were redissolved in 1 mL of 100% vol/vol LC-MS-grade methanol. A 500-μL aliquot of each extract was then transferred into a 96-well plate and used for LC-MS/MS analysis.
8
Residual Nitrite Analysis in Packaged Food
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Residual nitrite analysis was done in accordance with AOAC Method 973.31 (AOAC International, 2005b) . Sampling was conducted in duplicate on days 1, 6, 13, 27, 41, 55, 69, 83, 97, 111 , and 125 post packaging. Samples were prepared by separating the 2 exterior slices (slices 1 and 4 in the stack) from the 2 interior slices (slices 2 and 3 in the stack) of each package. The exterior and interior slices were finely chopped separately using a food processor (KitchenAid, St. Joseph, MI), and 5.0 g (± 0.01 g) was weighed into a beaker. Each 5.0 g sample was then added to a 500 mL volumetric flask with approximately 300 mL hot (approximately 50°C-80°C) distilled water and placed into a 100°C water bath. Flasks remained in the water bath for 2 h and were swirled every 30 min. After heating, the flasks were cooled to room temperature (approximately 23°C), then filled to volume (500 mL) with distilled water. Approximately 30 mL was filtered through Whatman #1 filter paper (Whatman Grade 1, GE Health Care Life Sciences, Pittsburgh, PA) into a 50 mL volumetric flask. Next, reagents were added as described in the AOAC International procedure, flasks were filled to volume, and absorbance at 540 nm was recorded using a Beckman DU 640 spectrophotometer (Model 4320940; Beckman, Fullerton, CA).
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