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A rapid and convenient method to prepare DIG-labelled RNA probes for use in non-radioactive in situ hybridization.

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Gandrillon, Olivier | Solari, F. | Legrand, C. | Jurdic, P. | Samarut, J.

Edité par CCSD ; Elsevier -

We describe here the use of PCR-generated templates incorporating T3 polymerase sites in order to prepare digoxigenin (DIG)-labelled cRNA probes against any gene of known sequence. This method was applied to the preparation of probes specific for chicken glyceraldehyde-3-phosphate dehydrogenase messenger RNAs and we demonstrate that such probes can be used for in situ hybridization (ISH). This technique therefore represents a rapid and convenient means to prepare DIG-labelled cRNA probes for use in a non-radioactive ISH. It adds speed and convenience of probe preparation to the previously described advantages of non-radioactive detection techniques.

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