Journalartikel

Jahr der Veröffentlichung: 1988

Seiten: 22-31

Zeitschrift: Gene Analysis Techniques

Bandnummer: 5

Heftnummer: 2

ISSN: 0735-0651

DOI Link: https://doi.org/10.1016/0735-0651(88)90023-4

Verlag: Elsevier

Abstract: 
A convenient and rapid method for preparing soluble extracts from the nuclei of as few as 3 × 10⁷
mammalian cells (miniextract procedure) is described. By several
criteria, miniextracts are comparable to nuclear extracts prepared from
large numbers of cells by the conventional procedure. Miniextracts are
able to support efficient transcription of a variety of class II
promoters. In addition, DNase I footprinting and gel retardation assays
can be performed directly in miniextracts, enabling the detection of
sequence-specific DNA-binding proteins. Besides transcription,
miniextracts efficiently carry out pre-mRNA splicing and allow formation
and fractionation of previously characterized splicing complexes. The
small-scale procedure enables simultaneous preparation of multiple
extracts from a variety of cell types under different experimental
conditions. Moreover, the use of small amounts of cells allows minimal
expenditure of valuable or expensive materials such as radioactive
compounds. Consequently, the procedure is highly advantageous for
biochemical analysis of transcription and RNA processing in mammalian
cells.

Harvard-Zitierstil: Lee, K., Bindereif, A. and Green, M. (1988) A small-scale procedure for preparation of nuclear extracts that support efficient transcription and pre-mRNA splicing, Gene Analysis Techniques, 5(2), pp. 22-31. https://doi.org/10.1016/0735-0651(88)90023-4

APA-Zitierstil: Lee, K., Bindereif, A., & Green, M. (1988). A small-scale procedure for preparation of nuclear extracts that support efficient transcription and pre-mRNA splicing. Gene Analysis Techniques. 5(2), 22-31. https://doi.org/10.1016/0735-0651(88)90023-4