Applications and improvements to the differential display method for gene analysis

Objective: Clone the differentially expressed genes between the well and poorly differentiated cholangiocarcinoma. Methods: A simple modification of primers and PCR conditions that gives rise to a more powerful mRNA differential display. A reverse Northern analysis that effectively eliminates the false positives isolated from differential display of mRNAs is applied. Preparation of probe by one-step labeling in reverse transcription reaction is found to be more effective and specific. Results: there are 9 fragments cloned. Conclusion: Analysis of multiple fragments of single slot blot, and requirement of RNA only in small amounts as compared to conventional Northern makes the protocol quick, effective, and economic.