Biotin-11-dUTP |
| カタログ番号GC50503 |
Biotin-11-dUTPは、dTTPの蛍光代替品であり、PCR、ニック翻訳、cDNA合成、ランダムプライマー標識あるいはプライマー伸長の間で、DNAを酵素的に非放射線標識する。
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は、dTTPの蛍光代替品であり、PCR、ニック翻訳、cDNA合成、ランダムプライマー標識あるいはプライマー伸長の間で、DNAを酵素的に非放射線標識する[1, 2]。Biotin-11-dUTPは、逆転写酵素、Taq DNAポリメラーゼ、phi29 DNAポリメラーゼ、エキソヌクレアーゼ非活性Klenow断片、Klenow断片とDNAポリメラーゼを通じてDNAに取り込まれる[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.
このプランはガイドのみを提供し、ご自分のニーズに合わせてご調整下さい。
1.溶液の調製
(1)作業溶液：この製品は、溶液の形として提供されている。
注：実際の条件または文献に合わせて、最適の作業濃度を調製し、またはご自分で文献を参考にし勾配濃度を設置してください。作業溶液は調製し、ただちに使用してください。
2.Biotin-11-dUTPでのDNAビオチン化標識[1]（文献より作成、参考まで）
(1)プライマーを使い、標的の遺伝子断片をPCR増幅する。
(2)37℃で、20μMのBiotin-11-dUTP、100μMのデオキシヌクレオシド三リン酸(dNTP)と0.6単位のKlenow断片で増幅産物を5分間培養する。
(3)取り込まれなかったBiotin-11-dUTPとdNTPは、エタノール沈殿で除去される。
(4)ビオチン化された遺伝子断片を25μLの殺菌水に再懸濁し、室温で、持続的に回転しながら、250μLのストレプトアビジン-アガロースビーズで1時間混合させた。
(5)洗浄と溶出画分のUV特徴よるDNAビオチン化の測定。260、200、240と289nm（ビオチンに）、そして280nm（ストレプトアビジンに）で特徴的なUV吸光度でDNAビオチン化を定量した。
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 |
| 溶解度 | 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 | 231.8 μL | 1.1592 mL | 2.3184 mL |
| 10 mM | 115.9 μL | 579.6 μL | 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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