Random labeled single-stranded RNA probes can be synthesized by in vitro transcription. The reaction is catalyzed by bacteriophage T7 RNA polymerase that incorporates labeled NTPs (mostly UTP) as substitute for their natural counterpart using linear, RNA probe-encoding DNA as template.
The optimal ratio of labeled NTP/NTP in terms of product yield and labeling efficiency depends on both the type of labeled NTP and final application. Individual optimization of labeled NTP/NTP ratio can easily be achieved with the single nucleotide format of our HighYield T7 RNA Labeling Kits. They offer maximum optimization flexibility in contrast to fixed NTP labeling mixes combined with high product yields. The combination of (poly)-HRP-conjugated Biotin/Digoxigenin detection reagents with AF 488 (Alexa Fluor® 488)-, AF546 (Alexa Fluor® 546)- or AF594 (Alexa Fluor® 594)-labeled tyramide further increases detection sensitivity of up to 100-fold.