Biotin-14-CTP is a CTP analog that contains biotin attached at the N4-position via a 14-atom linker. The amount of material provided is sufficient for up to 20 transcription reactions (1ug template) using T7 RNA polymerase.
Biotin-labeled RNA can be synthesized by SP6, T3, or T7 RNA polymerase and the appropriate template in the presence of ATP, GTP, UTP, and biotin- 14-CTP. Using a 1:1 mixture of biotin-14-CTP:CTP (keeping the total concentration the same as the other nucleotides) will result in a high yield of full length biotin-labeled RNA. Changing the ratio of biotin-14-CTP:CTP to 4:1 (increasing the total concentration of the biotin-14-CTP:CTP mixture to 2.5 times the concentration of each of the other nucleotides) will increase the amount of biotinylation, resulting in the generation of more sensitive probe. Use of the higher biotin-14-CTP:CTP ratio will, however, result in a lower total yield. The biotin-labeled RNA can be detected colorimetrically using Steptavidin-Alkaline Phosphatase Conjugate and NBT/BCIP or by chemiluminescence using streptavidin alkaline phosphatase and an appropriate chemiluminescent substrate.
Storage and Stability:
Store at -20 degrees C. Stable for at least 12 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
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