|
| Status |
Public on Feb 07, 2013 |
| Title |
5922012008_B |
| Sample type |
RNA |
| |
|
| Source name |
Mouse testis
|
| Organism |
Mus musculus |
| Characteristics |
strain: F1 generation of CBA x C57BL/6J
tissue: Mouse testis
time: 49days
genotype/variation: NSun2 KO
|
| Growth protocol |
Two lines of NSun2-/- mice were generated using the ES cell line D014D11 (German Gene Trap Consortium) and the mouse line Nsun2tm1a(EUCOMM)Wtsi (The Wellcome Trust Sanger Institute).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was isolated from testis using Trizol (Life Technologies) and cDNA synthesis was performed on 1µg of RNA using the Superscript III Reverse Trancriptase kit from Invitrogen using random hexamers
|
| Label |
Biotin
|
| Label protocol |
250 ng of total RNA was converted to complementary RNA (cRNA) target using the Illumina TotalPrep-96 Kit (Ambion 4397949). Total RNA was reverse transcribed and converted to double-stranded cDNA using a T7 promoter-Oligo(dT) primer and purified with magnetic oligo(dT) beads. This formed the template for an in vitro transcription (IVT) reaction which included a biotinylated nucleotide / ribonucleotide mix for both cRNA amplification and biotin labeling.
|
| |
|
| Hybridization protocol |
Standard Illlumina protocol (Illumina WGGX DirectHyb Assay Guide 11286331 RevA)
|
| Scan protocol |
Standard Illlumina protocol (Illumina WGGX DirectHyb Assay Guide 11286331 RevA)
|
| Description |
KO49E
|
| Data processing |
Using the R beadarray package the data was corrected for spatial artefacts, logged (base 2) and quantile normalised
|
| |
|
| Submission date |
Jul 18, 2012 |
| Last update date |
Feb 07, 2013 |
| Contact name |
Suraj Menon |
| E-mail(s) |
suraj.menon@cruk.cam.ac.uk
|
| Organization name |
Cambridge Research Institute, Cancer Research UK
|
| Street address |
Li Ka Shing Center, Robinson Way
|
| City |
Cambridge |
| ZIP/Postal code |
CB2 0RE |
| Country |
United Kingdom |
| |
|
| Platform ID |
GPL6887 |
| Series (1) |
| GSE39480 |
The mouse cytosine-5 RNA methyltransferase NSun2 is a component of the chromatoid body and required for testis differentiation |
|
