Liver_ASO_PND5_SMA_2
Mus musculus
Protocols: RNA purity, integrity and concentration were determined using an Agilent 2100 Bioanalyzer (Agilent Technologies, Inc, USA). Only RNA with a RIN value of 8.0 or higher and a ratio (28s/18s) of above 1.8 was taken forward for sample preparation. Samples were processed for library constructions following the manufactory instructions (Illumina Tru Seq Stranded Total RNA sample preparation v2 Guide, Part #15031048 Rev.E October 2013 - “Low sample protocol”)*. In brief, 0.5 µg of total RNA from each tissue were depleted for cytoplasmic rRNA using the Ribo-Zero ribosomal reduction chemistry, chemically fragmented for 8 min at 94oC, and processed for 1st strand synthesis cDNA and then 2nd strand synthesis using dUTP instead of dTTP. After some purification steps, the cDNA was then end-repaired, purified, adenylated at the 3'-ends, and purified before adding the indexed adaptor sequences using the TruSeq Stranded LT Kit Index set A. Each library preparation was then enriched by 10 cycles of PCR, purified and finally validated in regards to size and concentration. For sizing, libraries were analyzed on the Agilent 2100 Bioanalyzer using a DNA 1000 kit from Agilent Technologies. The Libraries were quantified by qPCR using the KaPa Library quantification Kits (KaPa Biosystems, Cat KK4824). Samples were pooled in sets of 12 libraries, and a final concentration of 16 pM denatured library was used for paired-end deep sequencing using an Illumina HiSeq1500.
ENA first public
2016-08-25
ENA last update
2016-07-25
External Id
SAMEA4355076
INSDC center alias
University of Southern Denmark Department of Biochemistry and Molecular Biology
INSDC center name
University of Southern Denmark Department of Biochemistry and Molecular Biology
INSDC first public
2016-08-25T17:01:19Z
INSDC last update
2016-07-25T12:22:57Z
INSDC status
public
Submitter Id
E-MTAB-36641469444294:Liver_ASO_PND5_SMA_2
age
5
broker name
ArrayExpress
common name
house mouse
genotype
spinal muscular atrophy
organism part
liver
sample name
E-MTAB-36641469444294:Liver_ASO_PND5_SMA_2
sex
male
treatment
2xSC100 MOE ASO