Preparation of mRNA and cDNA Libraries From Platelets and Megakaryocytes

The challenge of working with platelet and megakaryocyte mRNA is contamination with leukocyte mRNA. When attempting to detect mRNA encoding platelet or megakaryocytic specific proteins using the extremely sensitive technique reverse transcription polymerase chain reaction (RT-PCR); for either mRNA analysis or cloning, the concern for a few contaminating leukocytes is greatly amplified. Platelets pose an additional challenge because they contain only small amounts of residual mRNA from the parent megakaryocyte. Therefore, molecular biology techniques that require large amounts of RNA, such as Northern hybridization, have limited application. Usually, RT-PCR is used to answer questions of gene expression or to provide material for cDNA libraries for the cloning of proteins.