Drosophila Genetics Protocols

Chromatin Assembly on Template DNA with Transcription Factors and Drosophila S-190 Chromatin Assembl Protocol described here produces chromatin with regularly spaced nucleosomes having physiological nucleosome repeat lengths; something that can be difficult to achieve with purified components. In addition, chromatin assembled with the S-190 Chromatin Assembly Extract contains the ATP-dependent chromatin remodeling factors necessary for efficient transcription.

Detection of Even-Skipped Transcripts in Drosophila Embryos with PCR/DIG-Labeled DNA Probes Protocol for detection of even-skipped transcripts in Drosophila embryos with PCR/DIG-labeled DNA probes.

Drosophila Non-fluorescent Protein and RNA Double-Labeling Protocol Protocol for Drosophila non-fluorescent protein and RNA double-labeling.

Drosophila Chromatin Assembly Protocol (S150 extract) Protocol for Drosophila chromatin assembly (S150 extract).

FISH Protocols for Drosophila FISH protocols for Drosophila. Includes: RNA Probe Preparation; Embryo Collection and Fixation; Single FISH on Drosophila embryos; Post-Fixation, Hybridization and Post-Hybridization Washes; Development of FISH Signal; Storage, Mounting and Viewing of Samples; Double FISH on Drosophila Embryos; RNA-Protein Double Labeling; FISH on Dissected Tissues.

Nucleosomal Array Disruption Assay Protocol Protocol describes how to test whether a transcription factor disrupts the chromatin of a promoter of a gene of interest. First, chromatin is assembled in vitro on the gene of interest in the presence and absence of a transcriptional activator (see Protocol on Assembly of Chromatin with Drosophila S-190 Chromatin Assembly Extract and Transcriptional Activators).

Optimized Protocols for Fluorescent in situ Hybridization in Drosophila Tissues Optimized protocols for fluorescent in situ hybridization in Drosophila tissues. Includes: RNA Probe Preparation; Initial Embryo Fixation; Post-Fixation, Hybridization and post-Hybridization Washes; Development of FISH Signal; Mounting and Viewing of Samples; Double FISH; FISH on Dissected Tissues; RNA-Protein Double-labeling.

Preparation of a Highly Efficient Transcription Extract from Drosophila Embryos Protocol Protocol describes how to produce a soluble nuclear extract rich in basal pol II transcription factors from Drosophila embryos. This is a cell-free extract that contains all the necessary transcription factors and is capable of accurate initiation of transcription by RNA polymerase II but is deficient in core histones and histone H1.

RNA Affinity Isolations with PUMILIO from Drosophila melanogaster Protocol Protocol for RNA affinity isolation with PUMILIO from Drosophila melanogaster.

Whole Mount In Situ Hybridization for the Detection of mRNA in Drosophila Embryos Protocol Protocol for whole mount in situ hybridization for the detection of mRNA in Drosophila embryos. Includes: Embryo collection and fixation; Prehybridization treatment; Hybridization; Washing and detection.

X-Ray Mutagenesis of Drosophila Protocol Protocol describes the general procedure for creating mutations in the DNA of Drosophila by exposure to X-rays. Irradiation of cells with X-rays creates double strand breaks (DSBs) in DNA. Mutations introduced in the DNA of germ line cells (sperm) are propagated by mating the exposed males to virgin females. The progeny of this cross can be mated to each other so that a percentage of the subsequent offspring will have two copies of the same mutant allele.