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- W1036586781 abstract "Publisher Summary This chapter describes procedures for preparation and fractionation of yeast splicing extract. A two-step model has been formulated for pre-mRNA splicing. The first step involves cleavage at the 5′ splice site and the formation of a lariat structure. In the second step, the 3′ splice site is cleaved, and the two exons are joined. To understand the mechanism of the splicing reaction, it is necessary to identify components of the splicing machinery. The spheroplasts are collected by centrifugation at 3000 rpm for 5 min and washed once by gentle resuspension in 15 ml of SB3 buffer and then collected as before. The spheroplast pellets are gently resuspended in 8 ml of buffer A and lysed by homogenization in a Dounce homogenizer with strokes using a tight-fitting pestle. Pre-mRNA splicing reactions in the yeast whole cell extracts display rapid kinetics, unlike splicing in the mammalian system, in which there is a lag of 30–45 min." @default.
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- W1036586781 date "1990-01-01" @default.
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- W1036586781 title "Preparation and fractionation of yeast splicing extract" @default.
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- W1036586781 doi "https://doi.org/10.1016/0076-6879(90)81114-a" @default.
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