519789_2_PA_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). Small RNA library preparation was performed following the NEBNext Small RNA Library Prep Set for Illumina protocol. Briefly, 600 ng of total RNA was used to ligate 3' end Illumina-compatible adaptors, followed by hybridization with the reverse transcription primer that prevents formation of adaptor dimers; then 5' end adaptors were ligated to the small RNA, and adapter-ligated small RNA molecules were reverse transcribed and amplified by PCR with an initial denaturation at 94°C for 30 seconds, followed by 15 cycles of 94°C for 15 seconds, 62°C for 30 seconds and 70°C for 15 seconds, and a final extension step for 5 minutes at 70°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
SAMEA111422273
INSDC center alias
The Hospital for Sick Children
INSDC center name
The Hospital for Sick Children
INSDC first public
2023-05-31T00:15:34Z
INSDC last update
2023-05-31T00:15:34Z
INSDC status
public
Submitter Id
E-MTAB-12230:519789_2_PA_T
age
125
broker name
ArrayExpress
common name
sheep
developmental stage
embryo
disease
normal
genotype
wild type genotype
individual
519789_2
injury
none
isolate
not applicable
organism part
pulmonary artery
phenotype
wild type phenotype
sample name
E-MTAB-12230:519789_2_PA_T
scientific_name
Ovis aries
strain
Dorset x Rideau Arcott