2024-06-10
ENA first public
ERC000011
ENA-CHECKLIST
Gurdon Institute, University of Cambridge
INSDC center name
public
INSDC status
ERS20110431
SRA accession
E-MTAB-14104:BL17500 1
Submitter Id
ArrayExpress
broker name
not collected
collection date
fruit fly
common name
Protocols: In brief, groups of 50 testes for different genotypes were ground in 750 μl of TRIzol reagent and incubated at room temperature for 10 min.  Phenol was removed from samples by multiple rounds of chloroform extraction. RNA from the supernatant was precipitated by adding 0.5x isopropanol and washed once with 70% ethanol. The extracted RNA was then treated with RNase-free DNase I (New England Biolabs, M0303) for 30 min at 37°C to remove DNA followed by heat-inactivating DNase I by incubating 10 min at 65°C upon adding 1 μl of 50 mM EDTA.  Approximately 500 ng of total RNA was used for library prep. Following DNase I digestion, long RNA (>200 nt) was isolated using Zymo Clean and Concentrator columns (Zymo, R1019). Subsequently, rRNA was removed using the NEBNext mRNA Magnetic Isolation Module (New England Biolabs, E7490L). With the purified RNA, cDNA libraries were prepared by employing the NEBNext Ultra Directional RNA Library Prep Kit (New England Biolabs, E7760) and double-stranded cDNA was purified using AMPure XP Beads (Beckman Coulter, A63881).
description
y1 w67c23; P{EPgy2}Poldip2EY08866
genotype
not collected
geographic location (country and/or sea)
not applicable
isolate
Drosophila melanogaster
organism
Drosophila melanogaster
organism
testis
organism part
Drosophila melanogaster
scientific_name
BL17500 1
title