Glyco-gene microarray is a focused oligonucleotide microarray containing all of the cloned glyco-genes. It is high throughout technique to screen and detect the changes of transcription and expression levels of the glycan related genes such as glycan-binding proteins, glycosyltransferases, glycosylhydrolases, carbohydrate transporters, etc. It allows the analysis of RNA samples on a highly annotated custom gene microarray. The glyco-gene microarray is developed using Affymetrix technology. It contains probe sets to monitor the expression of approximately 2000 human and mouse transcripts and can also be custom-designed.
Figure 1. Summary of the glycan-related genes in human and mouse
The construction of glyco-gene microarray is similar to that of regular gene microarray. The representative glyco-genes compiled from currently available sequence database are selected and used for the microarray. Individual 59-mer oligonucleotides complementary to sequences selected are designed and prioritized using stringent selection criteria, including minimal secondary structure, minimal homology to other genes in the available human genomic databases, no low complexity or repeat regions, and all with a similar and well-defined Tm to provide optimal hybridization efficiency across all oligos on the micoarray. These optimal oligonucleotides are individually synthesized with the addition of a linker onto each oligonucleotide and then printed covalently linked in glass slides. The target RNA are extracted and labeled with fluorescence probes, followed by hybridization to each spot on the microarray. The results are quantitated by a high resolution confocal laser scanner. With glyco-gene microarray technology, it is possible to measure all of genes associated with glycoconjugate biosynthesis and degradation simultaneously.
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