Validating microarray data using rt real time pcr
Therefore, linear amplification protocols to increase the amount of RNA have been developed.
The correlation between the results of microarray experiments derived from non-amplified RNA and amplified samples needs to be evaluated in detail.
Approximately 80% of selected upregulated and downregulated genes identified by microarray analysis using linearly amplified RNA were confirmed by QRT-PCR using non-amplified m RNA as the starting template.
Conclusions: Linear RNA amplification methods can be used to generate high fidelity microarray expression data of comparable quality to data generated by microarray methods that use non-amplified m RNA samples.
Amplify your c DNA using Taq Man Assays & Applied Biosystems™ Taq Man® master mix.
However, standard hybridisation and detection protocols require micrograms of m RNA for microarray analysis, limiting broader application of this technology to small excisional biopsies, needle biopsies, and/or microdissected tissue samples.The use of real-time q PCR has nearly supplanted other approaches (e.g., Northern blotting, RNase protection assays). This review examines the current state of q PCR for gene expression analysis now that the method has reached a mature stage of development and implementation. A new quantitative PCR multiplex assay for rapid analysis of chromosome 17p11.2-12 duplications and deletions leading to HMSN/HNPP. Xiangqin Cui obtained her Ph D in Genetics in 2001 and did her postdoctoral training in Statistical Genetics from 2001 to 2004.She is currently an assistant professor in Department of Biostatistics, Section on Statistical Genetics, at the University of Alabama at Birmingham.