Problems in detecting CK19 mRNA with RT-PCR and corresponding countermeasures / 第二军医大学学报

ABSTRACT

Objective:To search for a better RT-PCR method for detecting CK19 mRNA. Methods: RT-PCR methods were used to detect the expression of CK19 mRNA in the tumor tissues and in the peripheral blood of patients with gastric cancer and in the peripheral blood of healthy subjects. Results: The influence of pseudogene and genome was observed through nonreversely transcripted RNA controls and comparison of several pairs of primers.The interference could be effectively eliminated by regulating the annealing temperature based on mismatched primers. Conclusion: The presence of pseudogene and genome interferes the experimental protocol in RT-PCR,which can be resolved with the method introduced in our paper.