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

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

doi:10.1016/0005-2787(77)90006-5

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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17 Scopus citations

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 language	English (US)
Pages (from-to)	228-237
Number of pages	10
Journal	BBA Section Nucleic Acids And Protein Synthesis
Volume	476
Issue number	3
DOIs	https://doi.org/10.1016/0005-2787(77)90006-5
State	Published - Jun 3 1977

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General Medicine

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10.1016/0005-2787(77)90006-5

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@article{30eda71fa225451cb582925bf151de08,

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

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.",

author = "Morrison, {Marcelle R.} and Fred Baskin and Rosenberg, {Roger N.}",

note = "Funding Information: Support for this work was providedi n part by a National Instituteso f Health, National Cancer InstituteP rojectGrant No. NIML-P01-CA17065-01 and by the AssociatedC ancerC enterC ore Tissue CultureF acilityand a grant from the Leland Fikes FoundationI,n c. The work in the paperw asc arriedo ut with the invaluablea ssistancoef Ms. Cathy Hall. Reticulocytpeo lysomeas nd globinm RNA samplews erea kind gift of ProfessoJre rry B. Lingrel.",

year = "1977",

month = jun,

day = "3",

doi = "10.1016/0005-2787(77)90006-5",

language = "English (US)",

volume = "476",

pages = "228--237",

journal = "BBA Section Nucleic Acids And Protein Synthesis",

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TY - JOUR

T1 - Quantitation and characterisation of poly(A)-containing messenger RNAs from mouse neuroblastoma cells

AU - Morrison, Marcelle R.

AU - Baskin, Fred

AU - Rosenberg, Roger N.

N1 - Funding Information: Support for this work was providedi n part by a National Instituteso f Health, National Cancer InstituteP rojectGrant No. NIML-P01-CA17065-01 and by the AssociatedC ancerC enterC ore Tissue CultureF acilityand a grant from the Leland Fikes FoundationI,n c. The work in the paperw asc arriedo ut with the invaluablea ssistancoef Ms. Cathy Hall. Reticulocytpeo lysomeas nd globinm RNA samplews erea kind gift of ProfessoJre rry B. Lingrel.

PY - 1977/6/3

Y1 - 1977/6/3

N2 - 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.

AB - 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.

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AN - SCOPUS:0017383501

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ER -

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

Abstract

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