A 60 µl solution of refolded gRNA 1–600 (NoProt condition) or of protein-RNA complex (Complex condition) was treated with 12 µl of 10 mM NMIA (Sigma-Aldrich) in anhydrous DMSO (Sigma-Aldrich) (+) or with 12 µl anhydrous DMSO (control, -). After 50 min at room temperature, 128 µl Milli-Q water were added and material was precipitated with 3 volumes of ethanol, 1/10 volume of 3 M sodium acetate pH 5.0, 1 µl of glycoBlue (ThermoFisher) for 30 min in a dry ice/ethanol bath and collected by centrifugation at 20,800 g for 30 min at 4°C. The pellets were washed twice with 70% ethanol, dried and resuspended in 15 µl water and 4 µl 5X Folding Buffer. One µl of proteinase K (Roche) was added and the digestion was performed at 37°C for 30 min before adding 79 µl water. The modified RNA was purified by phenol-chloroform extraction (1:1, pH 7.5) and ethanol precipitated as described above. RNA pellets were resuspended in 6 µl Milli-Q water.
Anhydrous dmso
Manufactured by Merck Group
Sourced in United States, Germany
Anhydrous DMSO is a colorless, odorless, and highly polar aprotic solvent commonly used in various laboratory applications. It has a high boiling point and low freezing point, making it a versatile solvent for a wide range of chemical reactions and procedures. Anhydrous DMSO is a valuable tool for researchers and scientists working in various scientific fields.
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Lab products found in correlation
Dimethyl sulfoxide (dmso), by Merck Group (5 mentions)
Nanodrop 2000c, by Thermo Fisher Scientific (4 mentions)
Neurobasal a medium, by Thermo Fisher Scientific (4 mentions)
Anhydrous dmf, by Merck Group (2 mentions)
Cm h2dcfda, by Thermo Fisher Scientific (2 mentions)
Cm h2dcfda, by Merck Group (2 mentions)
Basal media eagle media, by Thermo Fisher Scientific (2 mentions)
Alexa fluor 488 nhs ester, by Thermo Fisher Scientific (2 mentions)
Nap 5 column, by GE Healthcare (2 mentions)
High capacity rna to cdna master mix, by Thermo Fisher Scientific (2 mentions)
3 4 5 dimethyl 2 tiazolilo 2 5 diphenyl 2h tetrazolium bromide mtt, by Merck Group (2 mentions)
Dithiothreitol (dtt), by Merck Group (2 mentions)
Tri reagent, by Merck Group (2 mentions)
Streptomycin, by Merck Group (2 mentions)
Dnase 1, by Merck Group (2 mentions)
Hoechst 33342, by Thermo Fisher Scientific (2 mentions)
Alexa fluor 488, by Thermo Fisher Scientific (2 mentions)
Paclitaxel, by LC Laboratories (1 mentions)
Testosterone, by Merck Group (1 mentions)
Hexafluoro 2 propanol, by Merck Group (1 mentions)
56 protocols using anhydrous dmso
1
NMIA-Mediated RNA Structure Analysis
2
In vitro RNA structure probing
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Benzoyl cyanide (BzCN) probing was performed by splitting the sample into 25 μl aliquots and mixing with 2.78 μl of 400 mM BzCN (Sigma-Aldrich) dissolved in anhydrous DMSO (Sigma-Aldrich) [(+) sample)] or with anhydrous DMSO [(−) sample]29 (link),30 (link). Given that the BzCN probing reaction is complete within ~1 s, 75 μl of TRIzol LS reagent (Invitrogen) was added to each sample without any additional incubation time to stop the in vitro transcription reaction and the samples were vortexed.
Dimethyl sulfate (DMS) probing was performed by splitting the sample into 25 μl aliquots and mixing with 2.78 μl of 6.5% (v/v) DMS (Sigma-Aldrich) in anhydrous ethanol (Sigma-Aldrich) [(+) sample)] or with anhydrous ethanol [(−) sample] and incubating the samples at 37°C for 5 min. The DMS probing reaction was quenched by adding beta-mercaptoethanol to 2.8 M and incubating the sample at 37°C for 1 min. 75 μl of TRIzol LS reagent was added to each sample to stop the in vitro transcription reaction and the samples were vortexed.
Dimethyl sulfate (DMS) probing was performed by splitting the sample into 25 μl aliquots and mixing with 2.78 μl of 6.5% (v/v) DMS (Sigma-Aldrich) in anhydrous ethanol (Sigma-Aldrich) [(+) sample)] or with anhydrous ethanol [(−) sample] and incubating the samples at 37°C for 5 min. The DMS probing reaction was quenched by adding beta-mercaptoethanol to 2.8 M and incubating the sample at 37°C for 1 min. 75 μl of TRIzol LS reagent was added to each sample to stop the in vitro transcription reaction and the samples were vortexed.
3
Synthesis and Preparation of NAI
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NAI was synthesized as previously described (20 (link)). Briefly, 137.14 mg of 2-methylpyridine-3-carboxylic acid (Sigma Aldrich, cat. 325228) were resuspended in 500 μl DMSO anhydrous (Sigma Aldrich, cat. 276855). ∼162.15 mg of 1,1’-carbonyldiimidazole (Sigma Aldrich, cat. 115533) were resuspended in 500 μl DMSO anhydrous and added dropwise to the 2-methylpyridine-3-carboxylic acid solution while constantly stirring, over a period of 5 minutes. The reaction mixture was then incubated at room temperature with constant stirring for 2 h. This mixture (assumed to represent a 1 M stock) was aliquoted in 50 μl aliquots and stored at −80°C.
4
Preparation and Administration of Eribulin and Paclitaxel
5
Oligomeric Aβ Neurotoxicity and Testosterone Neuroprotection
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Oligomeric Aβ was prepared according to our previous report [31 (link)]. Aβ1–42 peptide was dissolved in 1,1,1,3,3,3-hexafluoro-2-propanol (Sigma) and then it was dried overnight by air at room temperature. The peptide pellet was resuspended with anhydrous DMSO (Sigma) at 2 mM as final concentration. Aβ was then bath-sonicated for 30 min at room temperature. Aliquot Aβ peptide was stored at −80°C deep freezer before use. The working concentration of Aβ was 5 μM in all experiments. This concentration of Aβ could induce synaptic degeneration including the reduction of synaptic proteins but not apoptosis when applied to hippocampal neurons for 24 h [28 (link), 31 (link)].
To investigate the neurotoxicity of Aβ on primary cultures of hippocampal neurons, Aβ was diluted with NB medium and was added to the cell culture at DIV14 for 24 h. Neuroprotective effects of testosterone were investigated in pretreatment experiments. 10 nM of testosterone has been considered to be at physiological dose [32 (link)]. Previous reports showed that 10 nM testosterone elicited neuroprotective effects in primary neuronal cultures [33 (link), 34 (link)]. Based on these findings, hippocampal neurons were incubated with 10 nM of testosterone (Sigma) for 1 h and then cotreated with oligomeric Aβ for 24 h. The control group was treated with DMSO as vehicle.
To investigate the neurotoxicity of Aβ on primary cultures of hippocampal neurons, Aβ was diluted with NB medium and was added to the cell culture at DIV14 for 24 h. Neuroprotective effects of testosterone were investigated in pretreatment experiments. 10 nM of testosterone has been considered to be at physiological dose [32 (link)]. Previous reports showed that 10 nM testosterone elicited neuroprotective effects in primary neuronal cultures [33 (link), 34 (link)]. Based on these findings, hippocampal neurons were incubated with 10 nM of testosterone (Sigma) for 1 h and then cotreated with oligomeric Aβ for 24 h. The control group was treated with DMSO as vehicle.
6
Synthesis and Characterization of Novel Polymers
7
Synthesis of Perovskite Solar Cell Materials
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Tin (IV) oxide (SnO2, 15% in H2O colloidal dispersion) was purchased
from Alfa Aesar. The MAPbI3 perovskite precursors, lead
(II) iodide (PbI2, 99%) and methylene ammonium iodide (MAI,
97%), were purchased from TCI. N2,N2,N2′,N2′,N7,N7,N7′,N7′-octakis
(4-methoxyphenyl)-9,9′- spirobi[9H-fluorene]-2,2′,7,7′
tetramine (spiro-OMeTAD, 99%) were all purchased from Borun Chem.
Exfoliated GO, dodecyl amine (DDA, ≥ 99%), glycine (≥99%),
anhydrous DMF (99.8%), anhydrous DMSO (≥ 99.5%), anhydrous
ethyl acetate (99.8%), anhydrous acetonitrile (99.8%), anhydrous toluene
(99.8%), anhydrous chlorobenzene (99.8%), and anhydrous hexane (95%)
were purchased from Sigma-Aldrich Company. All the chemicals and reagents
are directly used without any further purification.
from Alfa Aesar. The MAPbI3 perovskite precursors, lead
(II) iodide (PbI2, 99%) and methylene ammonium iodide (MAI,
97%), were purchased from TCI. N2,N2,N2′,N2′,N7,N7,N7′,N7′-octakis
(4-methoxyphenyl)-9,9′- spirobi[9H-fluorene]-2,2′,7,7′
tetramine (spiro-OMeTAD, 99%) were all purchased from Borun Chem.
Exfoliated GO, dodecyl amine (DDA, ≥ 99%), glycine (≥99%),
anhydrous DMF (99.8%), anhydrous DMSO (≥ 99.5%), anhydrous
ethyl acetate (99.8%), anhydrous acetonitrile (99.8%), anhydrous toluene
(99.8%), anhydrous chlorobenzene (99.8%), and anhydrous hexane (95%)
were purchased from Sigma-Aldrich Company. All the chemicals and reagents
are directly used without any further purification.
8
Synthesis and Purification of C*C Nucleotide
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0.3 mmol CMP (Sigma) was mixed with 0.15 mmol 2-amino-imidazole•HCl (Combi-blocks) in 5 ml anhydrous DMSO (Sigma) and 0.4 ml TEA (Sigma). Then 1 g triphenylphosphine (Sigma) and 0.88 g 2,2'-dipyridyldisulfide (Combi-blocks) were added in order and stirred vigorously. The reaction was continued in a sealed container for 20 min. The product C*C was precipitated by adding 40 ml acetone and 2 ml NaClO4-saturated acetone. After centrifugation at 3500 rpm for 10 min, the pellet was washed with 40 ml acetone:diethyl ether (1:1) and centrifuged again. The pellet was washed twice, then dried under house vacuum to remove organic solvent. The dry pellet was resuspended in 20 mM TEAB pH 8.0 and purified on a 50 g C18Aq column over 12 CV of 0–10% acetonitrile in 2 mM TEAB buffer (pH 8.0). The product was analyzed by 31P-NMR and low resolution mass spectrometry (LRMS) before being aliquoted and lyophilized.
9
Benzo(a)pyrene Exposure Protocol
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Anhydrous DMSO, corn oil, benzo(a)pyrene [B(a)P, purity 99%] and actinomycin D were from Sigma-Aldrich. Benzo(a)pyren-7, 8-dihydrodiol-9, 10-epoxide (BPDE, 99.9%) was from MRIGlobal (MRIGlobal). BPDE was dissolved in DMSO at and stored at . B(a)P was dissolved in corn oil before use. All the vehicle control or cultures included an equal amount of DMSO (0.03%, v/v).
10
Preparation of Oligomeric Amyloid-β Peptide
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