| Bioactivity | 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]. |
| Invitro | 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. |
| Name | Biotin-16-UTP |
| CAS | 186033-13-6 |
| Formula | C32H48Li4N7O19P3S |
| Molar Mass | 987.51 |
| Transport | Room temperature in continental US; may vary elsewhere. |
| Storage | Please store the product under the recommended conditions in the Certificate of Analysis. |
| Reference | [1]. G Patrone, et al. Nuclear run-on assay using biotin labeling, magnetic bead capture and analysis by fluorescence-based RT-PCR. Biotechniques. 2000 Nov;29(5):1012-4, 1016-7. |
Biotin-16-UTP
CAS: 186033-13-6 F: C32H48Li4N7O19P3S W: 987.51
Biotin-16-UTP is an active substrate for RNA polymerase.Biotin-16-UTP can replace UTP in the in vitro transcription reac
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