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3 protocols using agarose molecular grade

1

Comprehensive Chemical Reagents Protocol

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Methanol, isopropanol, dichloromethane, ethyl acetate, chloroform, and hexane were purchased from RCI Labscan Ltd., Thailand, and 95 % ethanol from J.T. Baker® (Center Valley, PA, USA). Sodium hydroxide, citric acid and hydrochloric acid were obtained from BDH Prolabo chemicals (Poole, UK) Neutral red and 4-(p-nitrobenzyl) pyridine (NBP) and acetonitrile were obtained from Sigma-Aldrich® (St. Louis, MO, USA). Melphalan, 4,6-diamidino-2-phenylindole (DAPI), copper (II) sulfate were from Sigma-Aldrich Chemie GmbH (Munich, Germany). Dulbecco’s modified Eagle medium (DMEM), RPMI Media 1640, fetal bovine serum and penicillin streptomycin were from GIBCO, Invitrogen Corp. (Grand Island, NY, USA). Sodium chloride and boric acid were from Vivantis Inc., (Oceanside, CA, USA). Dimethylsullfoxide (DMSO) (molecular grade) was from Sigma-Aldrich Chemie GmbH (Saint-Quentin Fallavier, France). Trypan blue was from Fluka Chemika (Buchs, Switzerland) and potassium sodium tartrate was from Loba Chemie (Mumbai, India). Agarose molecular grade was purchased from Bio-Rad (Hercules, CA, USA). A 100 bp DNA ladder marker was purchased from Invitrogen (Carlsbad, CA, USA). Caspase-Glo® 3/7, 8 and 9 were purchased from Promega (Madison, WI, USA).
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Molecular Biology Toolkit: Reagents Inventory

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Ethidium bromide (EtBr), ethylene glycol-bis (β-aminoethyl ether)-N,N,N′,N′-tetraacetic acid (EGTA), ethylenediaminetetra acetic acid (EDTA), isopropyl-β-D-thiogalactoside (IPTG), 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) coelenterazine, carbenicillin, CaCl2, Carbonyl cyanide m-chloro phenylhydrazone (CCCP), Calmidazolium (CDZ), Verapamil, coelenterazine, Trifluoperazine (TPZ), Chlorpromazine (CPZ), Phosphate Buffer Solution (PBS), Triton X-100, Tris-Acetate EDTA buffer (TAE), and aequorin oligonucleotides were all purchased from Sigma-Aldrich (St. Louis, MO, United States). Taq polymerase master mix for PCR (Promega, Madison, WI, United States), agarose molecular grade and 100 bp molecular ruler were obtained from Bio-Rad, Hercules, CA, United States). Kanamycin, Gentamicin, Streptomycin, Vancomycin, Ciprofloxacin, and Erythromycin (St. Louis, MO, United States). BamHI and HindIII (Rowley, MA, United States), T4 ligase (Rowley, MA, United States).
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3

Cytotoxicity Assay Protocol for Cell Line Studies

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The organic solvents used for extraction were of analytical grade and obtained from Fisher Scientific (Loughborough, UK) and Labscan (Bangkok, Thailand). The reagents used in the cell assays were of molecular biological grade. Dulbecco’s modified Eagle’s medium (DMEM), fetal bovine serum (FBS), penicillin, and streptomycin were purchased from GIBCO® (Invitrogen Corporation, Carlsbad, CA, USA). Standard reagents, melphalan, and 4′,6-diamidino-2-phenylindole (DAPI) were obtained from Sigma-Aldrich Inc. (St. Louis, MO, USA). Dimethylsulfoxide (DMSO) was purchased from United States Biological (Salem, MA, USA). The reagents for cytotoxicity testing included neutral red (NR) and sodium bicarbonate (NaHCO3). We used a FlexiGene DNA Kit (Qiagen GmbH, Germany), isopropyl alcohol biotechnology grade (Bio Basic Inc., USA), and methanol analytical grade (BDH, England). Agarose molecular grade was purchased from Bio-Rad (Hercules, CA, USA). We purchased a 100 bp + 1.5 kb DNA ladder stained with loading dye from SibEnzyme Ltd. (Russia). Caspase-Glo® 3/7, 8, and 9 Assay Kits were purchased from Promega (Madison, WI, USA). We obtained 3,3′-dihexyloxacarbocyanine iodide (DiOC6) from Sigma-Aldrich Inc. Pure concentrated hydrogen peroxide (35%, v/v) was purchased from Qrec Co. Ltd. (New Zealand).
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