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JBC, Vol. 252, Issue 14, 5040-5053, Jul, 1977
C. A. Marotta, J. T. Wilson, B. G. Forget and S. M. Weissman
Sequences of human beta-globin mRNA were determined by analysis of
complementary DNA. beta-mRNA was transcribed into double-stranded cDNA by
RNA-dependent DNA polymerase. cDNA was cut by restriction endonucleases and
the fragments were terminally labeled by means of polynucleotide kinase and
[gamma-32P]ATP. After purification, fragments were degraded by snake venom
phosphodiesterase. Alternatively single-stranded [32P]cDNA was prepared by
transcription in the presence of [alpha-32P]dCTP and actinomycin D; the
product was digested by endonuclease IV and degraded by snake venom
phosphodiesterase. cDNA tracts obtained by both labeling methods enabled us
to construct a sequence for the translated and 3'-terminal untranslated
regions of human beta-mRNA.
Human beta-globin messenger RNA. III. Nucleotide sequences derived from complementary DNA
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