AIMTo explore a method to replace the isotope probe in the PCR-Select differential screening Kit with DIG labeled probe.

METHODSDifferential expressed sequences isolated from Suppression Subtractive Hybridization (SSH) was translocated onto nitric fibrin film. Probes were prepared with DIG-11-dUTP mixed in by PCR. Hybridization and coloration followed routine operation procedures of DIG labeled probe. Positive hybridization results were verified with reverse Northern blot.

RESULTSThe positive results from the PCR-Select differential screening Kit improved with DIG labeled probe achieved 90% correspondence verified by reverse Northern blot.

CONCLUSIONThe PCR-Select differential screening Kit improved with DIG labeled probe maintained high specificity while avoiding isotope pollution. It can replace isotope probe completely.