100 free local cams - Validating microarray data with real time rt pcr
However, Northern blotting is used to assess large variations in expression levels and, thus, errors due to approximate normalization appear to be acceptable since they will not dramatically affect the derived trends in the expression of the target genes.
Microarray analysis has enabled gene expression analysis to be expanded from investigations of one gene at a time to wide-scale analyses, in which expression levels of many genes can be monitored simultaneously.
We found that real-time RT-PCR using Taq Man Low-Density Arrays yielded reproducible measurements over seven orders of magnitude.
Our model identified numerous genes that were expressed at nearly constant levels, including the housekeeping genes PGK1, GAPD, GUSB, TFRC, and 18S r RNA.
Internal standards, added to the starting material and quantified in parallel to the target analytes, are commonly used for this purpose.