[CAS NO. 186033-13-6] Biotin-16-UTP
PRODUCTS SPECIFICATIONS [186033-13-6]
DESCRIPTION [186033-13-6]
Overview
| MDL | - |
|---|---|
| Molecular Weight | 987.51 |
| Molecular Formula | C32H48Li4N7O19P3S |
| SMILES | [H][C@]12CS[C@@H](CCCCC(NCCCCCC(NCCCC(NC/C=C/C(C(N3)=O)=CN([C@H]4[C@H](O)[C@H](O)[C@H](O4)COP(OP(OP(O[Li])(O[Li])=O)(O[Li])=O)(O[Li])=O)C3=O)=O)=O)=O)[C@@]1([H])NC(N2)=O |
Summary
Biotin-16-UTP is an active substrate for RNA polymerase. Biotin-16-UTP can replace UTP in the in vitro transcription reaction for RNA labeling [1] .
In Vitro
Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs).
In Vitro RNA Synthesis and Purification:
1. Incubate the cells according to your normal protocol.
2. Add gently One volume of transcription buffer 2x [200 mM KCI, 20 mM Tris-HCI, pH 8.0, 5 mM MgCl2, 4 mM dithiothreitol (DTT), 4 mM each of ATP, GTP and CTP, 200 mM sucrose and 20% glycerol] to nuclei in ice, form mixture.
3. Add 8 μL biotin-16-UTP (from 10 mM tetralithium sal) to the mixture, which is incubated for 30 min at 29°C.
4. Add 6 μL 250 mM CaCl2, 6 μL RNase-free DNase I (10 U/μL) and incubating for 10 min at 29°C to stop reaction.
5. Perform RNA purification of both nuclear run-on and total RNA according to the manufacturer's instructions.
6. Resuspend RNA in 50 uL diethylpyrocarbonate (DEPC)-treated water.
7. Labeled RNA was captured by streptavidin-coated magnetic beads.
8. RNA-binding beads are then used for random hexamer primed reverse transcription.
MCE has not independently confirmed the accuracy of these methods. They are for reference only.
Shipping
Room temperature in continental US; may vary elsewhere.
Storage
Please store the product under the recommended conditions in the Certificate of Analysis.