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Synthesis of DNAs complementary to human ribosomal RNAs polyadenylated in vitro
Abstract:
Conditions are described under which poly(A) polymerase from Escherichia coli ribosomes will catalyse the addition of AMP residues onto the 3'-ends of human 18 S and 28 S ribosomal RNAs at an average rate of 40 AMP residues per 1000 nucleotides in 20 min. Single-stranded complementary DNAs (cDNAs) can then be transcribed from the polyadenylated RNAs with RNA-directed DNA polymerase from avian myeloblastosis virus. All of the sequences in the RNAs are represented in the cDNAs; measurements of the rates at which the cDNAs hybridize with their template RNAs showed that, when appropriate adjustments for differences in lengths and G + C contents of the reacting sequences are taken into account, the Rot 1/2 values of homologous RNA-cDNA hybridization reactions are directly proportional to the base-sequence complexity of the RNAs.