Blank Pallflex TX40 FDMS filters were supplied by Air Monitors (Gloucestershire, UK). These are made of borosilicate microfibres reinforced with woven glass cloth and bonded with polytetrafluoroethylene (5 mg cm−2, 47 mm diameter). The pH of solutions was measured by using a Mettler-Teledo (SevenGo™) pH meter. Suspensions were shaken in an end-over-end rotator inside an incubator (Stuart® SI500 shaking incubator from Barloworld Scientific Ltd., Staffordshire, UK). All glassware and plastic ware were soaked overnight in 10% HNO3 then rinsed three times with deionised water before use. All chemicals were of analytical grade. Bovine serum albumin, NaH2PO4, KCl, pancreatin (porcine), urea, CaCl2·2H2O and pepsin (porcine) were purchased form Merck (Poole, UK). Glucose, NaCl, Na2SO4, NH4Cl, NaHCO3 and NaOH were supplied by VWR International (Lutterworth, UK). Lysozyme, glutathione, MgCl2·6H2O, glucuronic acid, glucosamine hydrochloride, mucin (porcine), DPPC, hydrochloric acid (HCl) (36.5–38%) and nitric acid (HNO3) (≥69% Trace SELECT® for trace analysis) were obtained from Sigma Aldrich (Gillingham, Dorset, UK). Glycine was purchased from Fisher Scientific (Loughborough, UK). Multi-element standard stock solution (10 mg L−1 of As, Cd, Cr, Cu, Mn, Ni, Pb and Zn) and Fe standard stock solution (1003 mg L−1) were obtained from Qmx Laboratories (Essex, UK).
Nh4cl
Manufactured by Avantor
Sourced in United Kingdom
NH4Cl is a chemical compound that serves as a laboratory reagent. It is commonly used in various scientific applications, such as in the preparation of buffers, as an electrolyte, and in specific analytical procedures. NH4Cl's core function is to provide a source of ammonium ions and chloride ions for various experimental and analytical needs.
Automatically generated - may contain errors
Lab products found in correlation
Sodium chloride (nacl), by Avantor (5 mentions)
Na2hpo4, by Avantor (2 mentions)
Kh2po4, by Avantor (2 mentions)
Bovine mucin, by Merck Group (2 mentions)
Bovine serum albumin (bsa), by Avantor (2 mentions)
Tryptone, by Avantor (2 mentions)
Mucin, by Merck Group (2 mentions)
K2hpo4, by Thermo Fisher Scientific (2 mentions)
Mgcl2 7h2o, by Thermo Fisher Scientific (2 mentions)
Nh2 2co, by Avantor (2 mentions)
Cacl2, by Avantor (2 mentions)
Nahco3, by Thermo Fisher Scientific (2 mentions)
Potassium chloride (kcl), by Avantor (2 mentions)
Kh2po4, by Merck Group (2 mentions)
Glucosamine hydrochloride, by Merck Group (1 mentions)
Glucuronic acid, by Merck Group (1 mentions)
Na2so4, by Avantor (1 mentions)
Nahco3, by Avantor (1 mentions)
Glycine, by Thermo Fisher Scientific (1 mentions)
Mgcl2 6h2o, by Merck Group (1 mentions)
6 protocols using nh4cl
1
Preparation and Characterization of Simulated Lung Fluids
2
Semisolid Agar Plate Preparation
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The preparation of semisolid “swimming” agar plates was carried out according to Tremblay et al. [41] (link). M9-medium, consisting of the following salts Na2HPO4, KH2PO4, NaCl and NH4Cl (all purchased from VWR Germany) diluted in aqua dest., and the supplements MgSO4, CaCl2 and glucose-monohydrat were pipetted to 0.3% Bacto agar (BM, Becton) dissolved in aqua dest. The solution was then filled into petri dishes and dried under laminar flow. The semisolid agar plates were used on the same day as preparation. For the motility-tests overnight cultures of the different strains were diluted in fresh LB broth and incubated for additional 2 h at 37°C. 10 µl of this bacterial suspension was inoculated in the middle of a semisolid agar plate and incubated at 37°C for 48 h. Experiments were repeated three times and the reported values represent the average. Error bars reflect the standard deviations.
3
Standardized SARS-CoV-2 Inoculum Matrices
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Low and high inoculum levels were developed in three separate inoculum matrices at a target concentration of 3.5 and 7.0 log PFU on surfaces, respectively. Sterile 1× phosphate-buffered saline (PBS), pH 7.4, was prepared by adding 100 μL of diluted Φ6 stock (high, 10-fold dilution; low, 1,000-fold dilution) to 5 mL of PBS. Similarly, 100 μL of diluted Φ6 stock (high, 10-fold dilution; low, 1,000-fold dilution) was added to 5 mL of artificial saliva consisting of 1.54 mM KH2PO4 (Sigma-Aldrich), 2.46 mM K2HPO4 (Fisher Scientific, Loughborough, UK), 0.04 mg/L MgCl2·7H2O (Alfa Aesar, Ward Hill, MA), 0.11 g/L NH4Cl (VWR), 0.12 g/L (NH2)2CO (VWR), 0.13 g/L CaCl2 (VWR), 0.19 g/L KSCN (Acros Organics, Carlsbad, CA), 0.42 g/L NaHCO3 (Fisher Scientific), 0.88 g/L NaCl (VWR), 1.04 g/L KCl (VWR), and 3 g/L mucin (Sigma-Aldrich) at pH 7 (30 , 31 (link)). Lastly, the tripartite matrix (5 mL) was prepared as described in international standard ASTM E2197-17 (24 ) by combining 3.4 mL of PBS containing Φ6 stock (low and high) with a 1.6-mL solution consisting of 0.8 mg/mL bovine mucin (Sigma-Aldrich, St. Louis, MO), 2.5 mg/mL bovine serum albumin (VWR), and 3.5 mg/mL tryptone (VWR) to mimic fluids shed by infected individuals (1 (link), 3 (link), 24 , 32 (link)).
4
Artificial Saliva and Fecal Matrix Preparation
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The artificial saliva matrix was chosen to mimic cough or sneeze ejecta and prepared according to ASTM E2721-16. In brief, the artificial saliva matrix consisted of 0.21 g/L KH2PO4 (Sigma-Aldrich, St. Louis, MO), 0.43 g/L K2HPO4 (Fisher Scientific, Loughborough, UK), 0.04 mg/L MgCl2·7H2O (Alfa Aesar, Ward Hill, MA), 0.11 g/L NH4Cl (VWR), 0.12 g/L (NH2)2CO (VWR), 0.13 g/L CaCl2 (VWR), 0.19 g/L KSCN (Acros Organics, Carlsbad, CA), 0.42 g/L NaHCO3 (Fisher Scientific), 0.88 g/L NaCl (VWR), 1.04 g/L KCl (VWR), and 3 g/L mucin (Sigma-Aldrich) at pH 7 (ASTM International, 2016 ; Owen et al., 2021 (link)). The tripartite matrix was chosen to mimic fecal material shed by infected individuals and prepared as per international standard ASTM E2197-17 (ASTM International, 2017 ). In brief, the tripartite matrix consisted of 0.8 g/L bovine mucin (Sigma-Aldrich), 2.5 g/L bovine serum albumin (VWR), and 3.5 g/L tryptone (VWR) (ASTM International, 2017 ; Kasloff et al., 2021 (link); Riddell et al., 2020 (link); Sattar et al., 2003 (link)). Phi6 stock was added to each matrix to obtain inoculum levels of approximately 7 log PFU/mL.
5
Deuterium-labeled Biochemical Experiments
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glycerol-d8 (99% D) and deuterium
oxide (99.9% D) were bought from ChemSupport AS (Hommelvik, Norway).
For the experiments at D-lab and DEMAX, these chemicals were purchased
from Eurisotop and Sigma-Aldrich, respectively. K2HPO4, KH2PO4, Na2HPO4, NH4Cl, and glycerol were from VWR (Oslo, Norway). All
other chemicals and chemical competent cells were purchased from Sigma-Aldrich
(Merck Life Science AS, Oslo, Norway).
oxide (99.9% D) were bought from ChemSupport AS (Hommelvik, Norway).
For the experiments at D-lab and DEMAX, these chemicals were purchased
from Eurisotop and Sigma-Aldrich, respectively. K2HPO4, KH2PO4, Na2HPO4, NH4Cl, and glycerol were from VWR (Oslo, Norway). All
other chemicals and chemical competent cells were purchased from Sigma-Aldrich
(Merck Life Science AS, Oslo, Norway).
6
Minimal E. coli Growth Medium
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This medium is the M9 minimal medium described for E. coli K12 in [15] and [26] . It is based on glucose as a limiting carbon source, a phosphate buffer, salts and micronutrients. This medium contains, per liter: 5 g D-glucose (Acros), 1 g NH 4 Cl (VWR Prolabo), 0.5 g NaCl (VWR Prolabo), 6 g Na 2 HPO 4 anhydrous (Acros), 3 g KH 2 PO 4 (Fisher Scientific). The following components are then added separately from a stock solution: 2 mL of 1 M MgSO 4 • 7 H 2 O (Chem-Lab), 1 mL of 0.1 M CaCl 2 (Sigma), 0.3 mL of 1 mM thiamine HCl (Sigma) and 10 mL of trace element solution. This trace element solution contains, per liter:
and 0.18 g of CoCl 2 • 6 H 2 O (Riedel de Haën). This growth medium is sterilized according to the protocol in Nanchen et al. [26] . The different solutions containing glucose, salts and phosphate buffer are separately autoclaved (VWR Vapour Line) and mixed afterwards. Then, the separately sterilized micronutrient solution is added after cooling of this mixture. thiamine HCl is filter sterilized through a 0.2 µm pore size filter (TRP Filtermax) due to thermal instability, as well as the trace element solution, to prevent precipitation of any poorly soluble salts during or after autoclaving.
and 0.18 g of CoCl 2 • 6 H 2 O (Riedel de Haën). This growth medium is sterilized according to the protocol in Nanchen et al. [26] . The different solutions containing glucose, salts and phosphate buffer are separately autoclaved (VWR Vapour Line) and mixed afterwards. Then, the separately sterilized micronutrient solution is added after cooling of this mixture. thiamine HCl is filter sterilized through a 0.2 µm pore size filter (TRP Filtermax) due to thermal instability, as well as the trace element solution, to prevent precipitation of any poorly soluble salts during or after autoclaving.
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