A more reliable and reproducible way to create fluorescently or radioactively labeled cDNA
David T. Wong Heidi Davis Darryl Garrison Terry
Payette Matt Lundy
Kim Kuplent Mary A. Buchanan Joseph A. Sorge Rebecca L. Mullinax
Travis Ward Andrew Firmin
Nucleic acid microrrays, which contain hundreds or thousands of genes in a single format, are increasingly being used to identify differentially expressed genes. In this process, test and control mRNA are converted into cDNA probes in a reverse transcription reaction that incorporates labeled nucleotides. The test and control cDNA probes are then hybridized to arrays, the unhybridized probe is removed, and the hybridization is detected. Differences in the levels of hybridization signals of specific genes on the array correlate with differences in the abundance of those genes in the test and control mRNA used to prepare the probes.
We have developed the Microarray Labeling KitPP for use with gene microarrays1 to increase the reliability and reproducibility of identifying differentially expressed genes. The kit includes several control mRNA and two Control Microarrays designed to assess the mRNA quality and the sensitivity and specificity of the assay.
The Microarray Labeling Kit includes three artificially generated mRNA from Arabidopsis
thaliana genes. They are A. thaliana RUBISCO activase (rca),
chlorophyll a/b-binding protein, and ribulose-1,5-biphosphate carboxylase/oxygenase
large subunit (rbcL) genes. These genes were selected because they are involved
in plant-specific processes and do not have known human homologues. In addition,
they do not hybridize to membrane-arrayed human cDNA from 29 different tissues.
Therefore, cDNA probes generated from these genes are unlikely to hy