Affymetrix GeneChip Drosophila Genome [DrosGenome1]

Identification of FGF-dependent genes in the Drosophila tracheal system

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Transcriptome analysis of the targeted overexpression of branchless (UAS-bnl) using a tracheal driver (breathless-GAL4) in comparison to a wildtype reference at two different developmental stages (10-11h AEL and 17-19h AEL), and of a bnl-P1 mutant in comparison to a wildtype reference at 17-19h AEL.

genotype

Transcription profiling of targeted overexpression of branchless (UAS-bn1) at different developmental stages, and of a bn1-P1 mutant in Drosophila

Transcriptome analysis of the targeted overexpression of branchless (UAS-bnl) using a tracheal driver (breathless-GAL4) in comparison to a wildtype reference at two different developmental stages (10-11h AEL and 17-19h AEL), and of a bnl-P1 mutant in comparison to a wildtype reference at 17-19h AEL.


Affymetrix:Protocol:Hybridization-Ronald

Followed manufacturer's recommendations, used protocol EukGE-WS2v4 and one additional round of staining for the arrays.

P-AFFY-2

Labeling of samples was carried out according to the manufacturer’s standard protocol

P-AFFY-6

Title: Affymetrix CEL analysis. Description:

P-MEXP-21753

We studied the targeted overexpression of a branchless (bnl) transgene under the control of a tracheal driver (breathless, btl-GAL4) at two different time points, as well as bnl homozygous mutant embryos carrying the P1 insertion (bnl-P1) at one time point. Reference to this analysis were animals bearing the tracheal driver, not the effector. At 25 degress C, overexpression embryos were sampled at 10-11h AEL (experiment 1) and 17-19h AEL (experiment 2), mutant embryos were sampled at 17-19h AEL (experiment 3). Reference embryos were collected at respective time points.

P-MEXP-21754

For each biological replicate and experiment, we combined embryonic material as follows: 1) Several rounds of embryo collection for several days. 2) At each round, embryonic membranes were removed with bleach, embryos were rinsed thoroughly with water, and dried embryos were snap frozen in liquid nitrogen and stored at -80 degrees C. 3) At the day of the experiment, embryos from several rounds (5-10) of collection were combined for RNA extraction.

P-MEXP-21755

Total RNA was prepared from 100-200mg of embryos using the RNeasy Mini-Kit, including QiaShredder columns (both Qiagen).

Transcription profiling of targeted overexpression of branchless (UAS-bn1) at different developmental stages, and of a bn1-P1 mutant in Drosophila

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