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[Interference of human processed pseudogene and corresponding counter-measures in reverse transcriptase-polymerase
1Department of Thoracic Surgery, the First Affilliated Hospital of Chongqing University of Medical Sciences, Chongqing,400016 P.R.China.
Objective:
This study aimed to set up an effective method to find the interference of processed pseudogene and overcome it in RT-PCR.
Methods:
Using RT-PCR methods, the authors amplified the cytokeratin 19 cDNA or DNA sequence in human lung squamous cell carcinoma samples.
Results:
They could find the influence of pseudogene through performing genomic DNA and nonreversed transcribed RNA controls. By digesting the extracted RNA sample with RNase-free DNase or regulating the annealing temperature based on mismatched primers, they could effectively eliminate the interference.
Conclusion:
The presence of processed pseudogene usually interferes in the experimental protocol in RT-PCR. This problem can be resolved with the method aforementioned.
