Quantitation and characterisation of poly(A)-containing messenger RNAs from mouse neuroblastoma cells.
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 6.5-10(5) with a range from 2.2-10(5) to 1.53-10(6). 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.
D000818:Animals; D002460:Cell Line; D005914:Globins / Q000096:biosynthesis; D007700:Kinetics; D051379:Mice; D008970:Molecular Weight; D009447:Neuroblastoma / Q000032:analysis*; D010944:Plants / Q000378:metabolism; D011061:Poly A / Q000302:isolation & purification*; D011132:Polyribosomes / Q000032:analysis; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000302:isolation & purification* / Q000378:metabolism; D012335:RNA, Ribosomal / Q000032:analysis; D014908:Triticum / Q000378:metabolism
D012333:RNA, Messenger; D012335:RNA, Ribosomal; D011061:Poly A; D005914:Globins