Journal article

A small-scale procedure for preparation of nuclear extracts that support efficient transcription and pre-mRNA splicing


Authors listLee, KAW; Bindereif, A; Green, MR

Publication year1988

Pages22-31

JournalGene Analysis Techniques

Volume number5

Issue number2

ISSN0735-0651

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

PublisherElsevier


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.




Citation Styles

Harvard Citation styleLee, 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 Citation styleLee, 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


Last updated on 2025-21-05 at 15:34