This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for 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 and human embryonic stem (ES) cell populations and differentiated cell populations derived from mouse ES cells.
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