Quantitation and characterisation of poly(A)-containing messenger RNAs from mouse neuroblastoma cells

Marcelle R. Morrison, Fred Baskin, Roger N. Rosenberg

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Abstract

Comparison of several isolation procedures for neuroblastoma poly(A)-containing mRNAs shows that the highest percentage recovery of undegraded and biologically active messenger RNAs is obtained using proteinase K prior to phenol extraction. The messenger RNAs thus isolated comprise approximately 1.5% of the total ribosomal RNAs and have negligible contamination with 18 and 28 S RNAs. On denaturing polyacrylamide gels they have an average molecular weight of 6.5 · 105 with a range from 2.2 · 105 to 1.53 · 106. The messenger RNAs have an average poly(A) content of 154 nucleotides. They are highly active in wheat germ in vitro protein synthesizing systems, giving as much as 4.3 pmol [35S]methionine incorporation into total protein per mol of mRNA. This is almost as active as a control globin mRNA preparation.

Original languageEnglish (US)
Pages (from-to)228-237
Number of pages10
JournalBBA Section Nucleic Acids And Protein Synthesis
Volume476
Issue number3
DOIs
StatePublished - Jun 3 1977

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ASJC Scopus subject areas

  • Medicine(all)

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