Protocol

In Vitro Transcription Using Drosophila Nuclear Extracts

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. This protocol describes RNA polymerase II transcription in vitro using a Drosophila embryonic nuclear extract called soluble nuclear fraction (SNF). Transcription and RNA purification are followed by detection of the transcripts by primer extension with reverse transcriptase and a labeled oligonucleotide.