SP6 RNA Polymerase can transcribe RNA from cloned DNA templates that are downstream from an SP6 promoter. The synthesis can be performed with labeled NTPs to generate highly labeled RNA. Synthesized RNA can be used in many applications, including:
RNA or DNA blotting techniques
In situ hybridization
RNase protection studies: Transcripts synthesized by the enzyme are used as precursor RNA for studies on RNA splicing and processing.
Synthesis of capped RNA in vitro with addition of m7GpppG or m7GpppA in excess over GTP or ATP during the transcription reaction. The generated antisense RNA can be introduced into cells to suppress the expression of the corresponding genes.
The synthesis of antisense RNA probes
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With this system, homogeneously labeled single-stranded RNA can be generated. Transcripts can be non-radioactively labeled with biotin or DIG-11-UTP or radioactively labeled to high specific activity with [α-32P] or [α-35S] labeled nucleotides.
