342286_2_LV_T
Ovis aries
Protocols: Under maternal general anesthesia and after fetal echocardiography, fetal lambs were delivered by hysterotomy. The umbilical cord was clamped and the lamb was euthanized with IV sodium pentobarbital (BimedaMTC, Cambridge, Canada). A median sternotomy was used to harvest the heart, then samples were taken from the LV, RV, AAo and main PA. Tissue was snap frozen with liquid N2 and stored at -80°C. At gestational age 76 days, coils were implanted in the left heart under continuous ultrasound guidance (Philips HDI 5000), using a completely percutaneous approach under strict aseptic conditions. A 20G needle (6” Quincke; Beckton-Dickinson) was used to deliver platinum coils (0.018” IDC, Boston Scientific) above the mitral valve until the LV appeared underfilled and the ascending aortic flow decreased or became retrograde. RNA was extracted using the miRNeasy Mini Kit (QIAGEN). RNA-Seq library preparation was performed following the TruSeq Stranded Total RNA Library Preparation protocol (Illumina, USA) with the Ribo-Zero Gold rRNA removal kit. Briefly, 600 ng of total RNA was used as the input material and submitted to rRNA depletion using biotinylated beads that contain rRNA-targeting-specific oligos. rRNA-depleted RNA samples were fragmented into ranges of 200-300-bases for 4 minutes at 94°C and converted to double stranded cDNA, end-repaired and adenylated at the 3' to create an overhang A to allow for ligation of Illumina adapters with an overhang T. Library fragments were amplified under the following conditions: initial denaturation at 98°C for 10 seconds, followed by 12 cycles of 98°C for 10 seconds, 60°C for 30 seconds and 72°C for 30 seconds, and finally an extension step for 5 minutes at 72°C. At the amplification step, each sample was amplified with a different barcoded adapter to allow for multiplex sequencing.
ENA-FIRST-PUBLIC
2023-05-31
ENA-LAST-UPDATE
2023-05-31
External Id
SAMEA111509988
INSDC center alias
The Hospital for Sick Children
INSDC center name
The Hospital for Sick Children
INSDC first public
2023-05-31T00:15:35Z
INSDC last update
2023-05-31T00:15:35Z
INSDC status
public
Submitter Id
E-MTAB-12327:342286_2_LV_T
age
125
broker name
ArrayExpress
common name
sheep
developmental stage
embryo
disease
normal
genotype
wild type genotype
individual
342286_2
injury
control
isolate
not applicable
organism part
heart left ventricle
phenotype
normal
sample name
E-MTAB-12327:342286_2_LV_T
scientific_name
Ovis aries
strain
Dorset x Rideau Arcott