Complementary techniques: validation of gene expression data by quantitative real time PCR.
Microarray technology can be considered the most powerful tool for screening gene expression profiles of biological samples. After data mining, results need to be validated with highly reliable biotechniques allowing for precise quantitation of transcriptional abundance of identified genes. Quantitative real time PCR (qrt-PCR) technology has recently reached a level of sensitivity, accuracy and practical ease that support its use as a routine bioinstrumentation for gene level measurement. Currently, qrt-PCR is considered by most experts the most appropriate method to confirm or confute microarray-generated data. The knowledge of the biochemical principles underlying qrt-PCR as well as some related technical issues must be beard in mind when using this biotechnology.
Provenzano
Maurizio
M
Department of Oncological and Surgical Sciences, University of Padova, via Giustiniani 2, Italy.
Mocellin
Simone
S
D000199:Actins / Q000737:chemistry; D000818:Animals; D001709:Biotechnology / Q000379:methods*; D019295:Computational Biology; D003627:Data Interpretation, Statistical; D020869:Gene Expression Profiling / Q000379:methods*; D005786:Gene Expression Regulation; D006801:Humans; D008957:Models, Genetic; D020411:Oligonucleotide Array Sequence Analysis; D016133:Polymerase Chain Reaction / Q000379:methods; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D012984:Software
D000199:Actins; D012333:RNA, Messenger