Quantitative isolation of RNA from human platelets.
We have adapted the acid-guanidinium-phenol-chloroform extraction procedure of Chomczinsky and Sacchi to achieve efficient rapid recovery of total RNA from human platelets. Sufficient platelet RNA (20 micrograms of total RNA per 30 ml of whole blood) can be recovered from relatively small individual samples to perform Northern blot analysis on individual donors and detect the mRNAs for glycoproteins IIb1(GP IIb) and IIIa1(GP IIIa), 3.4 kb and 6.2 kb, respectively. Platelet GP IIb and GP IIIa mRNAs could also be reverse transcribed, and amplified in vitro by the polymerase chain reaction (PCR). Thus, our technique allows simultaneous Northern blotting and PCR, and therefore should be of great help to the characterization of inherited platelet disorders such as Glanzmann's thrombasthenia.
Djaffar
I
I
U 150 INSERM, Hôpital Lariboisière, Paris, France.
Vilette
D
D
Bray
P F
PF
Rosa
J P
JP
D001792:Blood Platelets / Q000502:physiology*; D015152:Blotting, Northern; D015342:DNA Probes; D004587:Electrophoresis, Agar Gel; D006801:Humans; D008214:Lymphocytes / Q000502:physiology; D010980:Platelet Membrane Glycoproteins / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods; D012313:RNA / Q000097:blood* / Q000235:genetics / Q000302:isolation & purification; D012333:RNA, Messenger / Q000097:blood* / Q000235:genetics / Q000302:isolation & purification
D015342:DNA Probes; D010980:Platelet Membrane Glycoproteins; D012333:RNA, Messenger; D012313:RNA