GSM1964553: ham_low_1; Sus scrofa; RNA-Seq
RNA was isolated using TRI Reagent (Applied Biosystems) according to the manufacturer’s protocol. Samples of muscles were homogenised with beads using a Bullet Blender 24 homogenizer (Next Advance). RNA was purified by a bead method (Agencourt RNAClean XP kit) and its quality and quantity was assessed by a Qubit Fluorometer (Invitrogen) and TapeStation 2200 system (RNA tapes, Agilent). Ribosomal RNA was removed from 5000 ng of total RNA using a Ribo-Zero Gold rRNA Removal Kit (Human/Mouse/Rat) (Epicentre). The absence of rRNA was verified on the TapeStation 2200. A TruSeq RNA Sample Preparation Kit v2 (Illumina) was used to prepare cDNA libraries from 100 ng aliquots of the RNA-depleted samples according to the manufacturer’s protocol. The cDNA samples were ligated with indexed adaptors in the order shown in Table 2. Finally, the libraries were amplified in 15 cycles of PCR and their quantity was estimated using the Qubit 2.0 Fluorometer and 2200 TapeStation D1000 tapes. The final concentration of the cDNA libraries was normalised to 10 nM, after which they were pooled . The libraries were diluted according to a cluster generation protocol and loaded into a v3 Illumina flowcell (16 samples per line with four technical replicates)
GEO Accession
GSM1964553