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

on derivatized glass slides. Fluorescent probes were synthesized from spleen and polyA+ RNAs and were used to screen the microarray.

The enlarged area 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.

Conclusion

For generation of PCR amplicons for microarray analyses, the Taq from the Eppendorf MasterTaq kit performs exceptionally well, even when used at one-half of the recommended concentration. High yields of specific amplicons are obtained which can be used to generate high quality microarrays.

References

(1) Schena, M.; Shalon, D.; Davis, R.W.; and Brown, P.O. (1995). Quantitative monitoring of gene expression patterns with complementary DNA microarray. Science 270: 467-470.

(2) Lockhart, D.J.; Dong, H.; Byrne, M.C.; Follettie, M.T.; Gallo, M.V.; Chee, M.S.; Mittmann, M.; Wang, C.; Kobayashi, M.; Horton, H.; and Brown, E.L. (1996). Expression monitoring by hybridization to high-density oligonucleotide arrays. Nature Biotechnol. 14: 1675-1680.

(3) Hegde, P.; Rong Qi; Abernathy, K.; Gay, C.; Dharap, S.; Gaspard, R.; Earle-Hughes, J.; Snesrud, E.; Lee, N.; and Quackenbush, J. (2000). A Concise Guide to cDNA Microarray Analysis. Biotechniques 29: 548-562.









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