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Isolation of ribosomal RNA precursors from lung

✍ Scribed by James M. Hill

Publisher: Elsevier Science
Year: 1977
Tongue: English
Weight: 422 KB
Volume: 78
Category: Article
ISSN: 0003-2697
DOI: 10.1016/0003-2697(77)90096-3

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✦ Synopsis

A method is presented for the isolation of ribosomal RNA precursors from lung. The very labile precursors of ribosomal RNA in mouse lungs can be isolated and preserved when lung tissue (i) is frozen in liquid nitrogen and pulverized, (ii) is homogenized in hypotonic buffer, (iii) nuclei are rapidly isolated after being sieved through multiple screens, (iv) nuclei are treated with Tween-40 and Nadeoxycholate, (v) nuclei are digested with DNase, and (vi) partially purified nucleoli are pelleted through a sucrose gradient. Nucleolar RNA was extracted with phenol-SDS-chloroform.

The RNA was separated on polyacrylamide gels. Absorbance and radioacitve profiles of the RNAs on the gels can be obtained. Therefore, the specific activities of the ribosomal RNA precursors can be calculated.

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