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• W2299837860 endingPage "74" @default.
• W2299837860 startingPage "49" @default.
• W2299837860 abstract "The formation of most eukaryotic mRNA 3′ ends, referred to as polyadenylation, requires cleavage of a precursor followed by the sequential addition of adenylate residues. Nuclear extract is prepared from HeLa cells by a modification of the method developed for in vitro transcription. It is very similar to the protocol used to prepare mammalian splicing extracts. Lysis of the cells can be checked by examination under the light microscope. HeLa nuclei are much smaller than the unlysed cells, and by comparing aliquots taken before and after douncing, it is easy to tell if the cells have broken open. Extracts active for polyadenylation have also been made from lymphoid cells grown in tissue culture or harvested from the spleens of inoculated mice. For these cells, extraction of the nuclei with 300 mM NaC1 and one-third as much buffer C per cell gave better activity. Very recently, cleavage and polyadenylation activity has been detected in yeast whole cell extract." @default.
• W2299837860 created "2016-06-24" @default.
• W2299837860 creator A5029319648 @default.
• W2299837860 date "1990-01-01" @default.
• W2299837860 modified "2023-10-15" @default.
• W2299837860 title "Preparation of mammalian extracts active in polyadenylation" @default.
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• W2299837860 doi "https://doi.org/10.1016/0076-6879(90)81112-8" @default.
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