PCR-Based amplification of platelet mRNA sequences obtained from small-scale platelet samples

Platelet transcriptome studies provide a powerful tool in the analysis of disorders affecting the megakaryocytic-platelet lineage. However, individualised platelet gene expression profiling is hampered by the exceptionally low yield of platelet mRNA. The yield of mRNA transcripts that can be obtained from small-scale platelet preparations is generally not sufficient for standard RNA-labeling reactions used in expression profiling. Furthermore, leukocyte contaminants in platelet preparations are a potential source of 'unwanted' mRNA since they contain several orders of magnitude more mRNA than platelets. To overcome these limitations a strategy that combines leukocyte filtration and a PCR-based amplification technique (SMART) has been developed and extensively evaluated.