Project description:Our aim was to identify which stand alone amplification protocol performed best in our lab when the starting amount of total RNA was limited to 50 ng (A typical amount of total RNA we obtained by laser capture microdissection of tissues from silique samples). Since the quality of extracted RNA is very often tissue dependent and will affect the amplification efficiency, we decided to use total RNA extracted from whole siliques; better representing tissues targeted by LCM. We set out to compare two basic methods of RNA amplification, based on IVT and PCR. Keywords: Comparison of target preparation regimes from limiting amounts of RNA
Project description:Gene expression profiling of immortalized human mesenchymal stem cells with hTERT/E6/E7 transfected MSCs. hTERT may change gene expression in MSCs. Goal was to determine the gene expressions of immortalized MSCs.
Project description:For more than a decade, microarrays have been a powerful and widely used tool to explore the transcriptome of biological systems. However, the amount of biological material from cell sorting or laser capture microdissection is much too small to perform microarray studies. To address this issue, RNA amplification methods have been developed to generate sufficient targets from picogram amounts of total RNA to perform microarray hybridisation. In this study, four commercial protocols for amplification of picograms amounts of input RNA for microarray expression profiling were evaluated and compared. The quantitative and qualitative performances of the methods were assessed. Microarrays were hybridised with the amplified targets and the amplification protocols were compared with respect to the quality of expression profiles, reproducibility within a concentration range of input RNA, and sensitivity.
Project description:Transcriptional profiling of human mesenchymal stem cells comparing normoxic MSCs cells with hypoxic MSCs cells. Hypoxia may inhibit senescence of MSCs during expansion. Goal was to determine the effects of hypoxia on global MSCs gene expression.
