Genomic DNA (1 μg) was denatured using 0.3 M NaOH for 10 min at 37 °C. The samples were then incubated at 50 °C for 16 h after adding hydroquinone (Sigma-Aldrich, St. Louis, Missouri, USA) and sodium bisulfate (Sigma-Aldrich). Genomic DNA was analyzed via MSP using primer sets located within a CpG-rich area in the CLDN7 promoter. PCR samples were then resolved by electrophoresis on a 1.5% agarose gel. For the BGS assay, DNA was purified, and a CpG-rich promoter region was amplified by PCR. The PCR products were purified and cloned into a PCR 2.1-TA cloning vector (Invitrogen). A minimum of six positive clones from each product were selected for sequencing. The detailed primer sequences are shown in Additional file 3 : Table S2.
Sodium bisulfate
Manufactured by Merck Group
Sourced in United States
Sodium bisulfate is a chemical compound with the formula NaHSO4. It is a white, crystalline solid that is soluble in water. Sodium bisulfate is commonly used as a pH adjuster and water treatment agent in various industrial and laboratory applications.
Automatically generated - may contain errors
Lab products found in correlation
Hydroquinone, by Merck Group (2 mentions)
Ethanol, by Merck Group (2 mentions)
Sulfuric acid, by Merck Group (2 mentions)
Sodium acetate, by Merck Group (2 mentions)
Pcr2.1 ta cloning vector, by Thermo Fisher Scientific (1 mentions)
Sodium acetate trihydrate, by Merck Group (1 mentions)
Sodium thiosulfate solution, by Merck Group (1 mentions)
O dianisidine, by Merck Group (1 mentions)
Emsure iso, by Merck Group (1 mentions)
Sodium metabisulfite, by Merck Group (1 mentions)
Ammonium hydroxide, by Merck Group (1 mentions)
D fructose, by Merck Group (1 mentions)
Hydrochloric acid (hcl), by Merck Group (1 mentions)
Sucrose, by Merck Group (1 mentions)
D glucose, by Merck Group (1 mentions)
Sodium nitrate, by Merck Group (1 mentions)
Potassium hexacyanoferrate 2 trihydrate, by Merck Group (1 mentions)
Zinc acetate dehydrate, by Merck Group (1 mentions)
Starch, by Merck Group (1 mentions)
L arginine, by Merck Group (1 mentions)
8 protocols using sodium bisulfate
1
Methylation Analysis of CLDN7 Promoter
2
Comprehensive Chemical Reagents for Analytical Protocols
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The following chemicals were supplied from Sigma-Aldrich Co. (St. Louis, MO, USA): hydrochloric acid, ammonium hydroxide (28–30% v/v), ethanol (95% v/v), sodium nitrate (≥98% w/w), sulfuric acid (20% v/v), sodium thiosulfate solution (0.10 N), sodium acetate buffer pH 5.30, potassium hexacyanoferrate(II) trihydrate, sodium acetate trihydrate (≥98.5% w/w), zinc acetate dehydrate (≥98% w/w), sodium bisulfate, sodium metabisulfite (≥97.0% w/w), D-(−)-fructose, sucrose, D-(+)-glucose, peroxidase from horseradish type II, and o-dianisidine (peroxidase substrate). From Merck (Germany), the following were obtained: potassium iodide for analysis EMSURE® ISO, Reagent Ph Eur, starch (soluble guaranteed reagent for analysis), phosphate buffer pH = 6.50, and hydrogen peroxide (30% v/v). Sodium chloride was supplied from Penta (Czech Republic), glycerol standard reference from Hanna Instruments (Gunstock, Rhode Island, USA), and acetonitrile Chromasolv™, for HPLC, gradient grade (≥99.9%) from Honeywell, Riedel-de Haën.
Ultrapure water of HPLC grade from an ultrapure water purification system with resistance of 12–18 μΩ-cm was used throughout.
Ultrapure water of HPLC grade from an ultrapure water purification system with resistance of 12–18 μΩ-cm was used throughout.
3
Preparation and Characterization of Levodopa-Arginine Salt
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Example 7
Levodopa-Arginine salt was prepared as follows:
Levodopa (LD) [Teva] was weighed in a suitable container with L-arginine [Merck] (at molar ratio of 1:1.8) and a 0.2% (sodium bisulfate [Sigma] solution in water was added to obtain a final concentration of 4.4% L-Dopa. The mixture was heated to 65+10° C. with constant stirring. When the solids were completely dissolved, solution was filtered using 0.45 μM nylon membrane. The filtered solution was immediately frozen in dry ice and subsequently subjected to lyophilization. The filtered solution was immediately frozen in dry ice and subsequently subjected to lyophilzation. Off-white crystals were obtained and subsequently subjected to MS analysis. The MS analytical results (shown in
4
Methylation Analysis of CLDN6 Promoter
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Genomic DNA was extracted from MCF-7 cells using Wizard Genomic DNA Purification Kit (Promega), and 1 μg DNA was denatured by 0.3 M NaOH for 10 min at 37 °C. All samples were incubated at 50 °C for 16 h after adding hydroquinone (Sigma, USA) and sodium bisulfate (Sigma, USA). Methylation-specific PCR (MSP) was performed to analyze the methylation status at CpG-rich area of CLDN6 promoter. Primers specific for unmethylated DNA and methylated DNA were summarized in Table 1 . PCR was carried out for 30 cycles at 95 °C for 30s, 58 °C for 30s, followed by a final extension at 72 °C for 10 min. PCR products were electrophoresed on 1.5 % agarose gel stained with ethidium bromide.
5
Starch-based Polymer Synthesis Protocol
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Starch (Analar BHD chemie Ltd., Poole, UK), acetic acid (East Anglia Chemicals, Fakenham, UK), potassium persulphate (≥99.0%—Sigma-Aldrich, St. Louis, MO, USA), sodium bisulfate (99.0%—Sigma-Aldrich), diethylene triamine 99% (Sigma Aldrich), maleic anhydrate (99%—Acros Organics, Somerville, NJ, USA), sodium acetate 99% (Sigma Aldrich), ethanol absolute 99% (Fisher Chemical, Waltham, MA, USA), and analytical grade chemicals and solvents were used in all the experiments.
6
Detailed Chemical Analysis Protocol
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Sodium bisulfate, copper (ii) sulfate, potassium sodium tartrate tetrahydrate, tannic acid, Folin-Ciocalteu, 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt, sodium acetate, HEPES, minimum 99.5% titration, 2,4,6-tris (2-pyridyl)-5-triazine (TPTZ), methyl viologen dichloride hydrate (Paraquat), amyloglucosidase from Aspergillus niger, and 2,2-di(4-tert-octylphenyl)-1-picrylhydrazyl free radical (DPPH) were purchased from Sigma-Aldrich (São Paulo, SP, Brazil). Glucose PAP liquiform (Kit) was purchased from Labtest (Belo Horizonte, MG, BRA) and potassium ferrocyanide trihydrate, zinc acetate dihydrate, iron(II) ammonium sulfate hexahydrate, gallic acid monohydrate, aluminum chloride hexahydrate, potassium persulfate, and iron(III) chloride were purchased from Vetec Química Fina LTDA (Rio de Janeiro, RJ, BRA). All the other chemicals used in this work were of analytical grade.
7
Arsenic Removal Protocol using Carboxymethyl Cellulose
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The BDH Chemicals Pvt. Ltd, Poole, UK supplied the sodium salt of carboxymethyl cellulose (CMC) (2% solution 6–8 pH, low viscosity, the viscosity of a 1% solution in water at 20 °C 30–70c/s) was supplied by arsenic trioxide (As2O3), sodium bisulfate, sodium nitrite, sodium hydroxide, and ethanol were supplied by Sigma-Aldrich Chemie Pvt, Ltd, USA. 0.1 mM of As(iii ) stock solution was prepared in double distilled water, and 0.1 M phosphate buffer solutions (PBS) in the varying pH range (5.7, 6.5, 7.0.7.5, and 8.0 pH) were made by adjusting the ratio of Na2HPO4 and NaH2PO4. All other chemicals were bought from Sigma-Aldrich Chemie Pvt, Ltd USA and utilized as received.
8
Synthesis of 1-Butylimidazole from Imidazole
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Imidazole (Him), methyl Imidazole (mim), butyl bromide, 1,4butanesultone, sodium ethoxide, p-nitrophenyl acetate, sulfuric acid (97-98%), and sodium bisulfate used as precursors or reactants were from Sigma-Aldrich. Methanol, sulfuric ether, ethyl acetate, hexane, acetone, butyl alcohol, and dichloromethane used as solvents or eluents were from Sigma-Aldrich. 1-ButylImidazole (bim) was synthesized by reaction of Imidazole with butyl bromide through an aliphatic nucleophilic substitution, following a previously reported method that employs sodium ethoxide for deprotonating the Imidazole, 27, 28 as it described in the ESI. †
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