Biotin-16-UTP tetrasodium

Guide (The following is the experimental plan we recommend. This plan serves only as a reference guide. The specific operations should be adjusted according to your actual needs.)
Extracellular RNA Synthesis and Purification [1] : Cells were cultured according to the conventional experimental protocol.
2. Gently add an equal volume of transcription buffer 2× (200 mM KCl, 20 mM Tris-HCl, pH 8.0, 5 mM MgCl?, 4 mM dithiothreitol (DTT), 4 mM each of ATP, GTP and CTP, 200 mM sucrose and 20% glycerol) to the cell nucleus, and mix thoroughly.
3. Add 8 μL of 10 mM Biotin-16-UTP tetrasodium to the mixture and incubate at 29°C for 30 minutes.
4. Add 6 μL of 250 mM CaCl? and 6 μL of RNase-free DNase I (10 U/μL), and incubate at 29°C for 10 minutes to terminate the reaction.
5. Purify the intranuclear extended RNA and total RNA.
6. Resuspend the RNA in 50 μL of water that has been treated with diethylpyrocarbonate (DEPC).
7. Use magnetic beads coated with streptavidin to capture the labeled RNA.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.