A RAPID AND CONVENIENT METHOD TO PREPARE DIG-LABELED RNA PROBES FOR USE IN NONRADIOACTIVE IN-SITU HYBRIDIZATION

We describe here the use of PCR-generated templates incorporating T3 p olymerase sites in order to prepare digoxigenin (DIG)-labelled cRNA pr obes against any gene of known sequence. This method was applied to th e preparation of probes specific for chicken glyceraldehyde-3-phosphat e dehydrogenase messenger RNAs and we demonstrate that such probes can be used for in situ hybridization (ISH). This technique therefore rep resents a rapid and convenient means to prepare DIG-labelled cRNA prob es for use in a non-radioactive ISH. It adds speed and convenience of probe preparation to the previously described advantages of non-radioa ctive detection techniques. (C) 1996 Academic Press Limited