title: GSM4802433: Heart of V3599K_1 without TAC; Mus musculus; RNA-Seq
library_construction_protocol: Hearts were removed, and total RNA was isolated using RNeasy mini kit (Invitrogen). Polyadenylated  mRNA was purified with oligo dT beads (NEBNext Poly (A) mRNA magnet Isolation Module, New England Biolabs, NEB). Complementary DNA (cDNA) libraries for Illumina sequencing were generated with NEBNext Ultra II RNA library Prep kit (NEB) and NEBNextplex Oligos for Illumina. Fragmentation was performed with NEBNext Random Primers in NEBNext First Strand Synthesis Reaction Buffer at 94°C for 15 min, and reverse transcription was performed with NEBNext Strand Synthesis Enzyme Mix. After ligation, library fragments were enriched, and index sequences were inserted with PCR amplification. Following purification with AMPureXP beads (Beckman Coulter), the quality and the concentration of the libraries was evaluated using an Agilent 2200 TapeStation (D1000, Agilent Thermo Fisher). The confirmed libraries were mixed to equal molecular amounts against the clustering and sequencing on an Illumina Next-seq DNA sequencer with 75 bp pair-end cycle sequencing kit (Illumina).