Biotin-11-dUTP |
| رقم الكتالوجGC50503 |
Biotin-11-dUTP هو بديل الفلورسنت لـ dTTP
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 86303-25-5
Sample solution is provided at 25 µL, 10mM.
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Biotin-11-dUTP is a fluorescent alternative to dTTP that enzymatically non-radioactively labels DNA during PCR, nick translation, cDNA synthesis, random primer labeling, or primer extension[1, 2]. Biotin-11-dUTP can be incorporated into DNA by reverse transcriptase, Taq DNA polymerase, phi29 DNA polymerase, exonuclease-inactive Klenow fragment, Klenow fragment, and DNA polymerase[3, 4, 5].
References:
[1] Green M R, Sambrook J. Preparation of labeled DNA, RNA, and oligonucleotide probes[J]. Cold Spring Harbor Protocols, 2022, 2022(1): pdb. top100578.
[2] Stutika C, Gogol-Döring A, Botschen L, et al. A comprehensive RNA sequencing analysis of the adeno-associated virus (AAV) type 2 transcriptome reveals novel AAV transcripts, splice variants, and derived proteins[J]. Journal of virology, 2016, 90(3): 1278-1289.
[3] Steck A L. Oligonucleotide-modified Nuclotides[J]. 2013.
[4] Bharati B K, Swetha R K, Chatterji D. Identification and characterization of starvation induced msdgc-1 promoter involved in the c-di-GMP turnover[J]. Gene, 2013, 528(2): 99-108.
[5] Anderson J P, Angerer B, Loeb L A. Incorporation of reporter-labeled nucleotides by DNA polymerases[J]. Biotechniques, 2005, 38(2): 257-264.
This plan only provides a guide, please modify it to meet your specific needs.
1. Solution preparation
(1) Working solution: This product is provided in the form of a solution.
Note: Please adjust the optimal working concentration according to the actual situation or refer to the literature to set the gradient concentration by yourself. The working solution must be prepared and used immediately.
2. DNA biotinylation labeling with Biotin-11-dUTP[1] (from the literature, for reference only)
(1) PCR amplify the target gene fragment using primers.
(2) Incubate the amplicon with 20μM Biotin-11-dUTP, 100μM deoxynucleoside triphosphate (dNTP) and 0.6 units of Klenow fragment at 37°C for 5min.
(3) Unincorporated Biotin-11-dUTP and dNTP are removed by ethanol precipitation.
(4) Resuspend the biotinylated gene fragment in 25μL sterile water and mix with 250μL streptavidin-agarose beads at room temperature for 1h with continuous rotation.
(5) Measurement of DNA biotinylation by UV characteristics of wash and elution fractions. DNA biotinylation was quantified by characteristic UV absorbance at 260, 200, 240, and 289nm (for biotin) and at 280nm (for streptavidin).
References:
[1] Bharati B K, Swetha R K, Chatterji D. Identification and characterization of starvation induced msdgc-1 promoter involved in the c-di-GMP turnover[J]. Gene, 2013, 528(2): 99-108.
| Cas No. | 86303-25-5 | SDF | |
| Canonical SMILES | [H][C@]12CS[C@H]([C@]1(NC(N2)=O)[H])CCCCC(NCCCCCC(NC/C=C/C3=CN(C(NC3=O)=O)[C@H]4C[C@@H]([C@H](O4)COP([O-])(OP([O-])(OP([O-])([O-])=O)=O)=O)O)=O)=O.CC[N+]([H])(CC)CC.CC[N+]([H])(CC)CC.CC[N+]([H])(CC)CC.CC[N+]([H])(CC)CC | ||
| Formula | C28H45N6O17P3S | M.Wt | 862.68 |
| الذوبان | 10mM Tris-HCl buffer (supplied pre-dissolved - 1mM) to 1 mM | Storage | Store at -20°C |
| General tips | Please select the appropriate solvent to prepare the stock solution according to the
solubility of the product in different solvents; once the solution is prepared, please store it in
separate packages to avoid product failure caused by repeated freezing and thawing.Storage method
and period of the stock solution: When stored at -80°C, please use it within 6 months; when stored
at -20°C, please use it within 1 month. To increase solubility, heat the tube to 37°C and then oscillate in an ultrasonic bath for some time. |
||
| Shipping Condition | Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request. | ||
| Prepare stock solution | |||
|
1 mg | 5 mg | 10 mg |
| 1 mM | 1.1592 mL | 5.7959 mL | 11.5918 mL |
| 5 mM | 0.2318 mL | 1.1592 mL | 2.3184 mL |
| 10 mM | 0.1159 mL | 0.5796 mL | 1.1592 mL |
Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)
g
μL
Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)
Calculation results:
Working concentration: mg/ml;
Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )
Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O, mix and clarify.
Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.
Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such as vortex, ultrasound or hot water bath can be used to aid dissolving.
3. All of the above co-solvents are available for purchase on the GlpBio website.
Quality Control & SDS
- View current batch:
- Purity: >98.50%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Average Rating: 5 (Based on Reviews and 29 reference(s) in Google Scholar.)
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