Cross-platform comparison of SYBR Green real-time PCR with TaqMan PCR, microarrays and other gene expression measurement technologies evaluated in the MicroArray Quality Control (MAQC) study.
BACKGROUND
The MicroArray Quality Control (MAQC) project evaluated the inter- and intra-platform reproducibility of seven microarray platforms and three quantitative gene expression assays in profiling the expression of two commercially available Reference RNA samples (Nat Biotechnol 24:1115-22, 2006). The tested microarrays were the platforms from Affymetrix, Agilent Technologies, Applied Biosystems, GE Healthcare, Illumina, Eppendorf and the National Cancer Institute, and quantitative gene expression assays included TaqMan Gene Expression PCR Assay, Standardized (Sta) RT-PCRtrade mark and QuantiGene. The data showed great consistency in gene expression measurements across different microarray platforms, different technologies and test sites. However, SYBR Green real-time PCR, another common technique utilized by half of all real-time PCR users for gene expression measurement, was not addressed in the MAQC study. In the present study, we compared the performance of SYBR Green PCR with TaqMan PCR, microarrays and other quantitative technologies using the same two Reference RNA samples as the MAQC project. We assessed SYBR Green real-time PCR using commercially available RT2 Profilertrade mark PCR Arrays from SuperArray, containing primer pairs that have been experimentally validated to ensure gene-specificity and high amplification efficiency.


RESULTS
The SYBR Green PCR Arrays exhibit good reproducibility among different users, PCR instruments and test sites. In addition, the SYBR Green PCR Arrays have the highest concordance with TaqMan PCR, and a high level of concordance with other quantitative methods and microarrays that were evaluated in this study in terms of fold-change correlation and overlap of lists of differentially expressed genes.


CONCLUSIONS
These data demonstrate that SYBR Green real-time PCR delivers highly comparable results in gene expression measurement with TaqMan PCR and other high-density microarrays.
Arikawa
Emi
E
SuperArray Bioscience Corporation, Frederick, MD 21704, USA. earikawa@superarray.net
Sun
Yanyang
Y
Wang
Jie
J
Zhou
Qiong
Q
Ning
Baitang
B
Dial
Stacey L
SL
Guo
Lei
L
Yang
Jingping
J
D052160:Benzothiazoles; D003959:Diamines; D020869:Gene Expression Profiling / Q000379:methods* / Q000592:standards*; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods* / Q000592:standards*; D009930:Organic Chemicals / Q000737:chemistry; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000592:standards*; D011786:Quality Control; D011804:Quinolines; D012313:RNA / Q000032:analysis; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D052160:Benzothiazoles; D003959:Diamines; D009930:Organic Chemicals; D011804:Quinolines; C098022:SYBR Green I; D012313:RNA