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General Information
Name
RNA-Seq_mE5955_CME-W1-Cl.8+
Species
D. melanogaster
Assay type
FlyBase ID
FBlc0001312
Project
Data Provider
Title
RNA-Seq of D. melanogaster, CME-W1-Cl.8+ cell line, modENCODE, unstranded.
Biosample Source
Overview
Strain
Stage
Sex
Tissue isolated
Other tissues studied
Cell component
Key genes
Methods
Sample preparation

Cells were grown in M3 (FBrf0021731) supplemented with 2% fetal calf serum and human insulin 5 µg/ml, 2% fly extract (FBrf0104877).

Cells were maintained between 2E6 and 1E7 cells/mL, and were harvested at about 5E6 cells/mL.

Biosamples used (1)
Biosample
Type
Title
D. melanogaster, CME-W1-Cl.8+ cell line, source for RNA.
Assay Details
Key genes
Protocol

Total RNA was isolated using Trizol, then Qiagen RNeasy spin columns. mRNA was isolated using Oligotex kit, then fragmented using divalent cations under elevated temperature. First and second strand cDNA synthesis was 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):37

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_mE5955_CME-W1-Cl.8+
Name Synonyms
RNA-Seq of D. melanogaster, CME-W1-Cl.8+ cell line, modENCODE, unstranded.
Secondary FlyBase IDs
    References (2)