[Validation of the libraries of the serial analysis of gene expression by the application of real-time quantitative polymerase chain reaction].
OBJECTIVE
To validate and supplement the libraries of serial analysis of gene expression (SAGE) by the application of the real-time quantitative polymerase chain reaction PCR).


METHODS
The primers were designed based on the full sequences of the genes. Nine single matched tags, 6 multiple matched tags, 1 non-matched tag due to the update of the National Center for Biotechnology Information (NCBI) database, and 2 non-matched tags were selected to fulfill the validation of real-time PCR.


RESULTS
The genes were all specifically amplified by the primers pairs. The expressions of the single matched tags were identical to those of the SAGE libraries; however, the expressions of only 3 genes of the 6 multi-matched tags were identical to those of the SAGE libraries. The PCR data of the non-matched tag due to the update of the NCBI database were opposite to those of the SAGE libraries. The data did not support the significant difference of the non-matched gene of the SAGE libraries.


CONCLUSIONS
Real-time PCR is a reliable tool for the validation of high through-put data such as SAGE. The reliability of data depends on the match of the tags of the SAGE libraries.
Department of Neurobiology, Shanghai Research Institute of Acupuncture and Meridian, Shanghai University of Traditional Chinese Medicine, Shanghai 200030, China.
Lei-miao
LM
Yin
Qing-hua
QH
Zhang
Yu
Y
Wang
Yu-dong
YD
Xu
Yong-qing
YQ
Yang
D020869:Gene Expression Profiling / Q000379:methods*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D060888:Real-Time Polymerase Chain Reaction