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.
Institute for Transplantation Diagnostics and Cellular Therapeutics, University of Düsseldorf, Germany.
Jutta M
JM
Rox
Jens
J
Müller
Bernd
B
Pötzsch
D000818:Animals; D001792:Blood Platelets / Q000166:cytology / Q000378:metabolism*; D001800:Blood Specimen Collection / Q000379:methods*; D020869:Gene Expression Profiling / Q000379:methods*; D006801:Humans; D047589:Leukocyte Reduction Procedures / Q000379:methods; D012333:RNA, Messenger / Q000096:biosynthesis*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*
D012333:RNA, Messenger