Preparation of Nuclear Extracts from Drosophila Embryos
This protocol was adapted from “Preparation of Nuclear Extracts from Drosophila Embryos and In Vitro Transcription Analysis,” Chapter 31, in Drosophila Protocols (eds. Sullivan et al.). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2000.INTRODUCTION
Drosophila nuclear extracts are often used in gene expression studies. They can be used directly as a source of transcription factors, or they can be fractionated, in some cases, to purified proteins. This protocol describes the preparation of a Drosophila embryonic nuclear extract called soluble nuclear fraction (SNF). It uses a low-salt (0.1 M) extraction and produces a relatively low amount of nonspecific DNA-binding proteins. SNF is highly active and is suitable for many in vitro transcription experiments.
