The chemical reagents for the synthesis of Ru(bpy)32+ and activated Ru(bpy)32+, including cis-bis(2,2′-bipyridine) dichlororuthenium (II), 2,2′-bipyridine-4,4′-dicarboxylic acid, N-hydroxysuccinimide (NHS), N,N′-dicyclohexylcarbodiimide (DCC), N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (EDC), sodium hexafluorophosphate, and sodium borate, were obtained from Alfa Aesar Co., Ltd. Streptavidin magnetic beads were synthesized by New England BioLabs. Diethylpyrocarbonate (DEPC)-treated water and RNase inhibitor were obtained from Takara Biotechnology (Dalian) Co., Ltd. PBS (20× solution) and reagents related to electrophoresis were purchased from Shanghai Sangon Biotechnology Co. Ltd. SYBR I and SYBR II were purchased from Invitrogen. Reagent-grade L-lysine, t-butyloxycarbonyl (Boc), trifluoroacetic acid (TFA), and glutathione (GSH) were purchased from Sigma-Aldrich and used without further purification except where noted. All oligonucleotides and probes synthesized in this work were purified by Invitrogen. The commercial kit for telomerase extraction was obtained from Millipore Co. Ltd.
Diethylpyrocarbonate depc treated water
Manufactured by Takara Bio
Sourced in United States
Diethylpyrocarbonate (DEPC)-treated water is a laboratory reagent used to inactivate RNase enzymes. It is prepared by treating water with DEPC, which reacts with and inactivates RNase. This DEPC-treated water can be used in various RNA-related applications to prevent RNA degradation.
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Lab products found in correlation
Streptavidin magnetic beads, by New England Biolabs (1 mentions)
Rnase inhibitor, by Takara Bio (1 mentions)
Glutathione (gsh), by Merck Group (1 mentions)
Sodium borate, by Thermo Fisher Scientific (1 mentions)
Sybr 1, by Thermo Fisher Scientific (1 mentions)
Cis bis 2 2 bipyridine dichlororuthenium 2 2 2 bipyridine 4 4 dicarboxylic acid, by Thermo Fisher Scientific (1 mentions)
Nebuffer 3, by New England Biolabs (1 mentions)
Recombinant rnase inhibitor, by Takara Bio (1 mentions)
Thermopol reaction buffer, by New England Biolabs (1 mentions)
Dna ladder marker, by Takara Bio (1 mentions)
Dntps mixture, by Takara Bio (1 mentions)
2 protocols using diethylpyrocarbonate depc treated water
1
Synthesis and Activation of Ru(bpy)3 Complexes
2
Preparation of RNase-Free EXPAR Solutions
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All DNA oligonucleotides (HPLC-grade) were synthesized by Sangon Biological Engineering Technology & Services Co., Ltd. (Shanghai, China). HPLC-purity miRNAs, dNTPs mixture and DNA ladder marker were purchased from TaKaRa Biotechnology Co. Ltd. (Dalian, China). The sequences of all DNA oligonucleotides and miRNAs used in this study are given in Table 1 . The nickase Nt.BstNBI and Vent (exo-) DNA polymerase, 10× NEBuffer 3.1 and 10× ThermoPol Reaction Buffer were purchased from New England Biolabs (NEB, Beverly, MA, USA). SYBR Green I was purchased from Invitrogen Biotechnology Co. Ltd. (Waltham, MA, USA). Diethylpyrocarbonate (DEPC)-treated water, and Recombinant RNase Inhibitor were purchased from TaKaRa Biotechnology Co. Ltd. All other chemicals (at least analytical grade) were obtained from Sigma-Aldrich (St. Louis, MO, USA). All DNA and miRNAs oligonucleotides sequences were diluted in 1× TE buffer (pH 8.0) to give stock solutions of 100 and 20 µM, respectively. To create and maintain an RNase-free environment, the tips and tubes are RNase-free and require pretreatment to inactivate RNase. All EXPAR solutions were prepared in DEPC-treated deionized water.
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