Pseudothymidine
Based on 1 Customer Validation
Pseudothymidine is a C-nucleoside analog of thymidine.
For research use only. We do not sell to patients.
- Purity: 99.44%
- CAS No.: 65358-15-8
- Formula: C10H14N2O5
- Molecular Weight:242.23
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Storage:Powder -20°C, 3 years , 4°C, 2 years ; In solvent -80°C, 6 months , -20°C, 1 month
Biological Activity
Pseudothymidine is a C-nucleoside analog of thymidine[1]. The calculated ΔΔG°50/mod is -0.5 kcal/mol, with a ΔTm/mod of 0.82°C. For the duplexes containing nine dA-T/ψT pairs, the ΔTm/mod is -0.9°C and a ΔΔG°50/mod is +1.1 kcal/mol. The modification of the duplex containing 12 consecutive dA-T/ψT base pairs produces a ΔTm/mod of -0.9°C and a ΔΔG°50/mod of +1.2 kcal/mol[2].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
Chemical Information
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CAS No. 65358-15-8
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Appearance Solid
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Molecular Weight 242.23
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Formula C10H14N2O5
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Color Light yellow to yellow
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SMILES
O=C(NC1=O)N(C)C=C1[C@H]2C[C@H](O)[C@@H](CO)O2
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Synonyms
5-Methyl-2'-Deoxypseudouridin
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Shipping
Room temperature in continental US; may vary elsewhere.
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Storage
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month
Solvent & Solubility
DMSO : 25 mg/mL (103.21 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month. When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month. When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
Select the appropriate dissolution method based on your experimental animal and administration route.
- For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
- To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for In Vivo experiments, it is recommended to prepare freshly and use it on the same day.
- The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.
Add each solvent one by one: 10% DMSO 40% PEG300 5% Tween-80 45% Saline
Solubility: ≥ 2.5 mg/mL (10.32 mM); Clear solution
This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).
Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.
Preparation of Saline: Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution.
Add each solvent one by one: 10% DMSO 90% (20% SBE-β-CD in Saline)
Solubility: ≥ 2.5 mg/mL (10.32 mM); Clear solution
This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).
Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 900 μL 20% SBE-β-CD in Saline, and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C, storage for one week): 2 g SBE-β-CD powder is dissolved in 10 mL Saline, completely dissolve until clear.
Please enter the basic information of animal experiments:
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Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
Please enter your animal formula composition:
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%DMSO +
Recommended: Keep the proportion of DMSO in working solution below 2% if your animal is weak.
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%+
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+%Tween-80 + +
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%Saline +
The co-solvents required include: DMSO, . All of co-solvents are available by MedChemExpress (MCE). , Tween 80. All of co-solvents are available by MedChemExpress (MCE).
Working solution concentration: 0.22 mg/mL
Method for preparing stock solution: mg drug dissolved in μL DMSO. Stock solution concentration: mg/mL.
1. Take μL DMSO stock solution;
2. Add μL .
μL , mix evenly;
3. Then add μL Tween 80, mix evenly;
4. Then add μL
Please ensure that the stock solution in the first step is dissolved to a clear state, and add co-solvents in sequence. You can use ultrasonic heating (ultrasonic cleaner, recommended frequency 20-40 kHz), vortexing, etc. to assist dissolution.
Protocol
Thermal DNA duplex denaturation studies are performed with templates containing up to twelve consecutive dA residues that are paired with its complement template containing consecutive T or Pseudothymidine (ψT) residues. Experiments are performed in a buffer (45 mM NaCl, 45 mM sodium citrate, pH 8.1, final vol. 1.5 mL) containing template and its complement (1.5 μM of each). Absorbance (260 nm) is monitored over a range of 25.0 to 90.0°C with a change in temperature of 0.5°C/min for five heating cycles. The initial heating cycle is discarded and the Tm is determined by averaging the temperatures of the remaining four cycles. The ΔTm between similar duplexes is calculated by subtracting the Tm of the duplex containing standard bases from the Tm of the duplex containing C-glycosides (including Pseudothymidine)[2].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
Purity & Documentation
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Data Sheet (277 KB)
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SDS (393 KB)
- English - EN (393 KB)
- Français - FR (393 KB)
- Deutsch - DE (393 KB)
- Norwegian - NO (393 KB)
- Español - ES (393 KB)
- Swedish - SV (393 KB)
- Italian - IT (393 KB)
- Portuguese - PT (393 KB)
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Handling Instructions (2659 KB)
References
[1]. S Lutz, et al. An in vitro screening technique for DNA polymerases that can incorporate modified nucleotides. Pseudo-thymidine as a substrate for thermostable polymerases. Nucleic Acids Res. 1999 Jul 1; 27(13): 2792-2798. [Content Brief]
[2]. Havemann SA, et al. Incorporation of multiple sequential pseudothymidines by DNA polymerases and their impact on DNA duplex structure. Nucleosides Nucleotides Nucleic Acids. 2008 Mar;27(3):261-78. [Content Brief]
Complete Stock Solution Preparation Table
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month. When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
| Optional Solvent | Concentration Solvent Mass | 1 mg | 5 mg | 10 mg | 25 mg |
|---|---|---|---|---|---|
| DMSO | 1 mM | 4.1283 mL | 20.6415 mL | 41.2829 mL | 103.2073 mL |
| 5 mM | 0.8257 mL | 4.1283 mL | 8.2566 mL | 20.6415 mL | |
| 10 mM | 0.4128 mL | 2.0641 mL | 4.1283 mL | 10.3207 mL | |
| 15 mM | 0.2752 mL | 1.3761 mL | 2.7522 mL | 6.8805 mL | |
| 20 mM | 0.2064 mL | 1.0321 mL | 2.0641 mL | 5.1604 mL | |
| 25 mM | 0.1651 mL | 0.8257 mL | 1.6513 mL | 4.1283 mL | |
| 30 mM | 0.1376 mL | 0.6880 mL | 1.3761 mL | 3.4402 mL | |
| 40 mM | 0.1032 mL | 0.5160 mL | 1.0321 mL | 2.5802 mL | |
| 50 mM | 0.0826 mL | 0.4128 mL | 0.8257 mL | 2.0641 mL | |
| 60 mM | 0.0688 mL | 0.3440 mL | 0.6880 mL | 1.7201 mL | |
| 80 mM | 0.0516 mL | 0.2580 mL | 0.5160 mL | 1.2901 mL | |
| 100 mM | 0.0413 mL | 0.2064 mL | 0.4128 mL | 1.0321 mL |