QRT-PCR TRANSCRIPT ANALYSIS

点击翻译按钮获取中文摘要

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. 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.