T7 RNA polymerase is commonly used to transcribe DNA which has been cloned into vectors which have two phage promoters in opposite orientation. RNA can be selectively synthesized from either strand of the insert DNA with different polymerases. Homogeneously labeled single-stranded RNA can be generated with this system. Transcripts can be non-radioactively labeled with biotin or DIG-11-UTP or radioactively labeled to high specific activity with [alpha-32P] or [alpha-35S]-labeled nucleotides.Synthesis of hybridization probes T7 RNA polymerase allows highly efficient production of homogeneously labeled RNA. This labeled RNA may be used as hybridization probes in Southern northern and dot blots as well as in situ hybridizations.Suitable labelsTranscripts can be nonradioactively labeled with biotin-16-UTP or DIG-11-UTP. They may also be radioactively labeled to high specific activity with [alpha-32P]- or [alpha-35S]-labeled nucleotides.