This subproject is one of many research subprojects utilizing theresources provided by a Center grant funded by NIH/NCRR. The subproject andinvestigator (PI) may have received primary funding from another NIH source,and thus could be represented in other CRISP entries. The institution listed isfor the Center, which is not necessarily the institution for the investigator.Transcript levels for various enzymes and proteins involved in murine cellular glycosylation and recognition are being quantitated by real-time quantitative RT-PCR (qRT-PCR) by a method that allows for medium-throughput (~800 genes) transcript analysis. A unified strategy for primer design is being combined with optimized strategies for mRNA isolation, DNA synthesis, and qRT-PCR that will allow for high efficiency detection and measurement of transcripts for glycan-related genes over a range of seven orders of magnitude in abundance. The approach is being applied for the measurement of 'glycan-related' transcripts from mouse embryonic stem (ES) cell populations and differentiated cell populations derived from mouse ES cells. In addition, the approach is being employed for the measurement of transcripts in mouse tissues and cell populations relevant to understanding the roles of glycan structures in animal physiology and pathology.
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