Enzymatic amplification of platelet-specific messenger RNA using the polymerase chain reaction.
Human platelets are derived from megakaryocytes as anucleate cells, and thus contain only vestigial amounts of RNA capable of being transcribed into protein. This has greatly hampered efforts to study directly platelet-specific gene products and their associated polymorphisms. In this report, we describe direct amplification, using the polymerase chain reaction, of platelet-derived mRNA in amounts sufficient to permit detailed analysis, such as restriction mapping and nucleotide sequencing. The ability to generate large amounts of cDNA from platelet-specific mRNA sequences should make possible direct molecular characterization of normal platelet proteins, and facilitate the investigation of a wide variety of inherited platelet disorders.
Newman
P J
PJ
Blood Center of Southeastern Wisconsin, Milwaukee 53233.
Gorski
J
J
White
G C
GC
Gidwitz
S
S
Cretney
C J
CJ
Aster
R H
RH
D001792:Blood Platelets / Q000201:enzymology* / Q000378:metabolism; D003001:Cloning, Molecular; D004247:DNA / Q000302:isolation & purification; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism*; D005784:Gene Amplification; D006801:Humans; D009693:Nucleic Acid Hybridization; D010980:Platelet Membrane Glycoproteins / Q000235:genetics; D012333:RNA, Messenger / Q000302:isolation & purification*
D010980:Platelet Membrane Glycoproteins; D012333:RNA, Messenger; D004247:DNA; D004259:DNA-Directed DNA Polymerase