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编 号：F747601
分子式：C32H48Li4N7O19P3S
分子量：987.51
分子式：C32H48Li4N7O19P3S
分子量：987.51
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CAS No: 186033-13-6
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生物活性：
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.
体外研究：
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.
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.
体外研究：
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.
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