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General Information
Name
RNA-Seq_mE4458_P6
Species
D. melanogaster
Assay type
FlyBase ID
FBlc0001337
Project
Data Provider
Title
RNA-Seq of D. melanogaster, iso-1 strain, pupa (24hr APF), modENCODE, unstranded.
Biosample Source
Overview
Sex
Other tissues studied
Cell component
Cell line
Key genes
Methods
Sample preparation

Fly stocks were reared in bottles at 24oC and after about 7 days, white prepupa on bottle walls were marked on the overlying glass and allowed to age as appropriate. Individuals were then transferred to a tube on dry ice.

Biosamples used (1)
Biosample
Type
Title
D. melanogaster, iso-1 strain, pupa (24hr APF), source for RNA.
Assay Details
Key genes
Protocol

Frozen samples were homogenized and extracted using the TRIzol reagent protocol (Invitrogen). RNA was purified on an RNeasy spin column (Qiagen), and DNase treated. Polyadenylated RNAs were purified from total RNA extracts via oligo(dT) binding, using standard Illumina protocol. The poly(A)+ RNA was fragmented using divalent cations under elevated temperature, following by first and second strand cDNA synthesis primed with random hexamers. The cDNA fragments were end-repaired using T4 DNA polymerase and Klenow DNA polymerase, and phosphorylated at their 5' ends with T4 polynucleotide kinase. After adding A bases to the 3' end of the DNA fragments, Illumina adaptor oligonucleotides were ligated to the ends and ~ 300 bp fragments were isolated from an agarose gel, enriched by PCR amplification, and gel-purified again.

Mode of Assay

Read length (bases):76

The samples were quantitated using a Nanodrop, and loaded onto a flow cell for cluster generation and sequenced on an Illumina Genome Analyzer II using either single read or paired end protocols (Illumina).

Comments
Associated Data
Size
Files
Additional Information
Synonyms and Secondary IDs (2)
Reported As
Symbol Synonym
RNA-Seq_mE4458_P6
Name Synonyms
RNA-Seq of D. melanogaster, iso-1 strain, pupa (24hr APF), modENCODE, unstranded.
Secondary FlyBase IDs
    References (7)