AGO1 IP and Drosophila Transcriptome

The samples were collected at 18°C, 25°C, or 29°C. AGO1 immunoprecipitation was performed as previously described (Kadener et al., 2009 (link); Lerner et al., 2015 (link)). Input and AGO1 IP Total RNA was extracted from fly heads using TRI reagent (Sigma) according to manufacturer’s protocol. cDNA synthesis was carried out as described in the Expression Analysis Technical Manual (Affymetrix). The cRNA reactions were carried out using the IVT kit (Affymetrix). Affymetrix high-density arrays for D. melanogaster version 2.0 were probed, hybridized, stained, and washed according to the manufacturer’s protocol.

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Martin Anduaga A., Evantal N., Patop I.L., Bartok O., Weiss R, & Kadener S. (2019). Thermosensitive alternative splicing senses and mediates temperature adaptation in Drosophila. eLife, 8, e44642.

Publication 2019

TRI reagent Expression Analysis Technical Manual

Cdna Crna Heads Immunoprecipitation Synthesis

Corresponding Organization :

Other organizations : Brandeis University, Hebrew University of Jerusalem

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Variable analysis

independent variables

Temperature (18°C, 25°C, or 29°C)

dependent variables

Gene expression levels determined by Affymetrix high-density arrays for D. melanogaster

control variables

AGO1 immunoprecipitation procedure, as previously described (Kadener et al., 2009; Lerner et al., 2015)
RNA extraction using TRI reagent
CDNA synthesis protocol
CRNA reaction using the IVT kit (Affymetrix)
Affymetrix high-density array hybridization, staining, and washing procedures

controls

Positive control: None specified
Negative control: None specified

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