GSM4816735: skin_5pm.B; Xiphophorus maculatus; RNA-Seq
Skin samples were dissected into separate 2.5 ml microcentrifuge tubes filled with RNAlater (Ambion Inc.) for RNA isolation.RNAlater was removed from respective microcentrifuge tubes followed by addition of 750 μL of QIAzol (Qiagen) in 2.0 mL collection tubes designed for automated tissue homogenization and RNA isolation stations. Skin were homogenized using the TisseLyser II (Qiagen) facilitated by stainless beads (Qiagen) for 10 min at 25 Hz. RNA isolation was subsequently performed using a QIAcube HT (Qiagen) automated bio-sample isolation system. The isolation system is equipped with a robotic arm with 8 pipettes. Each pipette is able to pick and eject pipette tips, self-clean, and transfers liquids between well/columns, or between reagent reservoirs and well/columns in standard 96-well plate formats. Each sample was independently maintained throughout the isolation process. Briefly, 150 µL of chloroform was added to each isolation tube and the samples were vigorously shaken for 15 sec and then phases partitioned by centrifugation (12,000×g for 15 min at 4°C).  The aqueous phase containing nucleic acids was transferred to a new sample tube by a rack of automated pipettors. After extraction, nucleic acids were precipitated with 500 µL 70% ethanol. RNA was then purified using a Qiagen RNeasy mini RNA kit (96-well plate) and eluted following the manufacturer's protocol. Each RNA sample was used to construct an independent sequencing library using Illumina TruSeq library preparation kit (Illumina, Inc., San Diego, CA, USA).
GEO Accession
GSM4816735