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Amplification of Mouse cDNAs for Microarrays Using the Eppendorf ,,, MasterTaq Kit

liver and spleen polyA+ RNAs

The final experiment performed was to verify that the PCR products amplified using the Eppendorf MasterTaq Kit generated strong specific signals in a microarray experiment. We printed 1,152 PCR products representing mouse cDNA clones and appropriate controls on a reflective slide (APBiotech, UK). Fluorescent probes were synthesized from mouse spleen and polyA+ RNAs and were used to screen the microarray (Fig. 3). Green, red, and yellow spots were observed, indicating positive hybridization of the labeled probes to the printed PCR products. The enlarged area shows the signal color range observed when using cyanine probes, Cye3 spleen, and Cye5 liver. Green (Cye3) and red (Cye5) spots indicate differential expression of these particular mRNAs sequences in spleen and liver respectively, whereas yellow spots indicate similar levels of expression in both tissues.

1152 cDNA clones derived from the mouse uniqene set and controls were amplified with the Eppendorf MasterTaq Kit and printed on derivatized glass slides. Fluorescent probes were synthesized from spleen and polyA+ RNAs and used to screen the microarray. The enlarged areas shows signal color range when using cyanine probes, Cye3 spleen and Cye5 liver. Green, red, and yellow spots indicate positive hybridization from the labeled probes. Green (Cye3) and red (Cye5) spots indicate that the targets are expressed differentially. Figure 3
The 1,152 cDNA clones derived from the mouse Unigene set and controls were amplified with the Eppendorf MasterTaq Kit and printed
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