Sample 14
Equus caballus
Protocols: Synovial fluid samples were obtained from both middle carpal joints prior to (day 0), day 28 and 70 days after surgery. Horses were sedated with detomidine (0.01 mg/kg, Domosedan Vet, Orion Pharma Animal Health, Copenhagen, Denmark) and butorphanol tartrate (0.01 mg/kg, Dolorex, MSD Animal Health, Copenhagen, Denmark), and the dorsal surface of the carpi prepared aseptically by chlorhexidine scrubbing and alcohol swabbing. Synovial fluid was aspirated using a 19 gauge 40 mm needle and immediately transferred into ethylene diamine tetra acetic acid (EDTA) tubes (BD Vacutainer, BD A/S, Albertslund, Denmark). Tubes were inverted carefully 5-10 times and samples were further processed as detailed in Table 1 within one hour and stored at -80 ‏⁰C until small RNA sequencing and RT-qPCR analysis of miRNA expression (performed within 6 months). 500 µl SF were treated with a 1ug/µl hyaluronidase (HA) solution to reduce viscosity as preciously described [12]. RNA was extracted from SF using miRNeasy serum/plasma Advanced Kit (Qiagen, Vedbæk, Denmark) according to the manufacturer's instructions. Samples underwent on-column DNase digestion with RNase free DNase sets (Qiagen, Vedbæk, Denmark) and RNA yield was determined by spectrophotometry (Nano Drop ND-1000 spectrophotometer, Saveen and Werner AB, Limhamn, Sweeden). This project utilized the NEBNext® Small RNA Library Prep Set for Illumina® (Multiplex Compatible). The protocol was adapted to accommodate the low input of sample by dilution of adapters 1 in 5 and extra PCR cycles.
ENA-FIRST-PUBLIC
2023-03-01
ENA-LAST-UPDATE
2023-03-01
External Id
SAMEA110141997
INSDC center alias
CGR, Institute of Infection Veterinary, and Ecological Sciences, University of Liverpool
INSDC center name
CGR, Institute of Infection Veterinary, and Ecological Sciences, University of Liverpool
INSDC first public
2023-03-01T00:16:44Z
INSDC last update
2023-03-01T00:16:44Z
INSDC status
public
Submitter Id
E-MTAB-11840:Sample 14
age
7
broker name
ArrayExpress
common name
horse
developmental stage
mature
individual
H6
organism part
synovial fluid
sample name
E-MTAB-11840:Sample 14
sampling site
midcarpal joint
scientific_name
Equus caballus
sex
male