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- W1455054676 abstract "Publisher Summary This chapter focuses on the isolation of amplified nucleoli from Xenopus oocytes. The chapter describes a special case in which a naturally occurring biological enrichment of a single gene makes it possible to isolate single genes as “native” chromatin. In the frog, Xenopus luevis , the DNA coding for the 18S and 28S ribosomal RNAs (rDNA) undergoes a massive amplification at the pachytene stage of oogenesis. This results in a nucleus that contains about 24 pgm of rDNA plus 12 pgm of bulk chromosomal DNA. By mid-diplotene stage, the amplified rDNA is packaged in several thousand extrachromosomal nucleoli, which are located close to the nuclear membrane. At this stage, the amplified nucleoli are actively engaged in ribosomal RNA synthesis and may be isolated completely free of other DNA contaminants. The first part of the procedure is designed to dissociate the ovary into single cells and to isolate those oocytes that have not yet begun to accumulate pigment or massive amounts of yolk. Later-stage oocytes are discarded because the large amounts of yolk and pigment that contain seriously interfere with nucleolar isolation. The immature oocytes are then homogenized, and nucleoli are isolated by banding them in buoyant-density gradients of metrizamide." @default.
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- W1455054676 date "1978-01-01" @default.
- W1455054676 modified "2023-10-03" @default.
- W1455054676 title "Chapter 4 Isolation of Amplified Nucleoli from Xenopus Oocytes" @default.
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- W1455054676 doi "https://doi.org/10.1016/s0091-679x(08)60132-6" @default.
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