title: Sample_46
organism: Equus caballus
description: Protocols: Percutaneous needle muscle biopsies (approx. 300mg) were obtained from the ventral compartment of the middle gluteal muscle from standing unsedated horses using a previously described method [53]and preserved in RNAlater (Thermo Fisher, Massachusetts, United States). Total RNA was extracted from approximately 70mg tissue, using a protocol combining TRIzol reagent (Thermo Fisher, Massachusetts, United States), DNase treatment (RNase free DNase) (Qiagen, Hilden, Germany) and RNeasy Mini-Kit (Qiagen). The study cohort comprised a subset of Thoroughbred horses, trained for Flat racing at a single training establishment under the management of a single trainer. The UR cohort comprised of yearling Thoroughbred horses (n = 51; 23 males and 28 females; 19.5±1.5 months old) in submaximal training prior to entering sprint training. The UE cohort consisted of the same group of horses undertaking their first or second 'work day' (WD, high-intensity sprint exercise simulating a race) on an all-weather gallop. The TR cohort consisted of horses at the end of the racing season following approximately six months of sprint training. All horses in this group had achieved “race fitness” with an average of 15.1±9.1 SD WDs (range 6-43). Table 1 summarises details of sampling and physiological measurements. Percutaneous needle muscle biopsies (approx. 300mg) were obtained from the ventral compartment of the middle gluteal muscle from standing unsedated horses using a previously described method [53]and preserved in RNAlater (Thermo Fisher, Massachusetts, United States). Total RNA was extracted from approximately 70mg tissue, using a protocol combining TRIzol reagent (Thermo Fisher, Massachusetts, United States), DNase treatment (RNase free DNase) (Qiagen, Hilden, Germany) and RNeasy Mini-Kit (Qiagen). RNA was quantified using a Nano Drop ND1000 spectrophotometer V 3.5.27 and RNA quality and purity were assessed using the 18S/28S ratio and RNA integrity number (RIN) on an Agilent Bioanalyser with the RNA 6000 Nano LabChip kit6 (Agilent, Cork, Ireland). The RNA isolated from UR, UE and TR samples had an average RIN of 8.1 (7.2-9.3), 8.2 (7-9.1) and 8.1 (5.4-8.6), respectively. Indexed, strand-specific Illumina sequencing libraries were prepared using the TruSeq Stranded mRNA Library Preparation Kit LT (Illumina, San Diego, United States).
attributes.tag: ENA first public
attributes.value: 2018-08-07
attributes.tag: ENA last update
attributes.value: 2017-01-30
attributes.tag: External Id
attributes.value: SAMEA59632918
attributes.tag: INSDC center alias
attributes.value: University College Dublin
attributes.tag: INSDC center name
attributes.value: University College Dublin
attributes.tag: INSDC first public
attributes.value: 2018-08-07T17:02:58Z
attributes.tag: INSDC last update
attributes.value: 2017-01-30T14:52:18Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-5447:Sample_46
attributes.tag: breed
attributes.value: Thoroughbred
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cohort
attributes.value: Trained_Rest
attributes.tag: common name
attributes.value: horse
attributes.tag: organism part
attributes.value: gluteus medius
attributes.tag: sample name
attributes.value: E-MTAB-5447:Sample_46