[Interference of human processed pseudogene and corresponding counter-measures in reverse transcriptase-polymerase chain reaction].
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.


CONCLUSIONS
The presence of processed pseudogene usually interferes in the experimental protocol in RT-PCR. This problem can be resolved with the method aforementioned.
Ge
M
M
Department of Thoracic Surgery, the First Affilliated Hospital of Chongqing University of Medical Sciences, Chongqing,400016 P.R.China.
Zhang
Y
Y
Li
L
L
Xi
B
B
Wang
M
M
Huang
T
T
D002294:Carcinoma, Squamous Cell / Q000235:genetics; D006801:Humans; D008175:Lung Neoplasms / Q000235:genetics; D016133:Polymerase Chain Reaction; D011544:Pseudogenes