Efficient RT-PCR on platelet mRNA after long-term storage.
We have developed a procedure permitting RT-PCR from mRNA even after a long-term storage (1 year) of platelet samples in ethanol (EtOH-platelets) at -80 degrees C. To validate our method, we have analysed the human platelet alloantigen system (HPA-1) which is coded by beta 3 mRNA. We have also demonstrated the efficiency of amplification of part of the coding region for (i) alpha IIb subunit mRNA, (ii) alpha v subunit mRNA, and (iii) the seven transmembrane domain thrombin receptor mRNA.
Peyruchaud
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O
UMR 5533 CNRS, Hôpital Cardiologique, Pessac, France.
Nurden
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A
Bourre
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F
D016824:Antigens, Human Platelet / Q000032:analysis*; D001792:Blood Platelets; D001793:Blood Preservation; D018955:CD36 Antigens / Q000032:analysis; D006801:Humans; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000032:analysis; D010980:Platelet Membrane Glycoproteins / Q000032:analysis*; D016133:Polymerase Chain Reaction / Q000379:methods*; D018807:Polymorphism, Single-Stranded Conformational; D012333:RNA, Messenger / Q000032:analysis*
D016824:Antigens, Human Platelet; D018955:CD36 Antigens; D019039:Platelet Glycoprotein GPIIb-IIIa Complex; D010980:Platelet Membrane Glycoproteins; D012333:RNA, Messenger