M Ge1, Y Zhang, L Li, B Xi, M Wang, T Huang. 1. Department of Thoracic Surgery, the First Affilliated Hospital of Chongqing University of Medical Sciences, Chongqing,400016 P.R.China.
Abstract
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.
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 humanlung 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.