Fco_09
Sus scrofa
Protocols: Ten different tissues from two unrelated one year old Landrace boars were included in the study. Hence, total RNA was extracted from heart, spleen, liver, kidney, lung, musculus longissimus dorsi, occipital cortex, hypothalamus, frontal cortex, and cerebellum employing the mirVanaTM RNA extraction kit (Ambion) according to manufacturers protocol, yielding a total of 20 samples. RNA integrity was assessed by manual inspection of the individual RNA samples on a 2% agarose gel. The RNA fragments were then copied into first strand cDNA using reverse transcriptase and random hexamer primers followed by second strand cDNA synthesis using DNA Polymerase I and RNaseH. The short cDNA fragments were finally prepared for sequencing on the Illumina Genome Analyzer using reagents provided in the Illumina Genomic DNA Sequencing Sample Prep Kit. The cDNA fragments were end-repaired using T4 DNA polymerase and Klenow DNA polymerase and a single adenine nucleotide base was added to the cDNA fragments by using 3 prime to 5 prime exonuclease. This was followed by ligation of Illumina adaptors, gel purification of fragments of approximately 200 bases in size and finally PCR enrichment for 15 cycles. The concentrations of the libraries were measured using Invitrogen Qubit fluorometer and the size and purity were determined using the Agilent DNA 1000 Kit on Agilent 2100 Bioanalyzer. The 89 libraries were diluted in buffer EB from QIAGEN to 10 nM, denaturated with 2 N NaOH to a final DNA concentration of 1.0 nM, diluted to 4 pM with pre-chilled hybridization buffer from Illumina and loaded in individual lanes into a 1.0 mm flowcell together with a single lane of a 2 pM PhiX control library from Illumina.
ENA-FIRST-PUBLIC
2013-02-20T17:01:55Z
ENA-LAST-UPDATE
2018-03-08T16:09:56Z
External Id
SAMEA2041071
INSDC center name
Aarhus University, Department of Molecular Biology and Genetics
INSDC first public
2013-02-20T17:01:55Z
INSDC last update
2018-03-08T16:09:56Z
INSDC status
public
Submitter Id
E-MTAB-1405:Fco_09
broker name
ArrayExpress
common name
pig
individual
147
organism part
frontal cortex
sample name
E-MTAB-1405:Fco_09
scientific_name
Sus scrofa
sex
male