GSM8758969: P21 Mouse retina RNA-seq Tet1/2/3 tcKO Replicate 3; Mus musculus; RNA-Seq
Retinas from P21 animals were dissected in PBS and homogenized in 1ml TRIZOL solution. 200µl of chloroform was added to homogenized TRIZOL solution, mixed by vortexing and incubated for 3 min at RT. Lysates were centrifuged for 10 min at 4C at max speed (16,000g). The aqueous phase with RNA was transferred into a fresh tube without disturbing the interphase. SMARTer Stranded Total RNA High Input (RiboGone Mammalian) (Takara) was used. Briefly, extracted RNA first undergoes ribosomal RNA removal. Then, we proceeded with cDNA synthesis and purification. Following, RNA-seq library was amplified by PCR. Total RNA was captured and purified from RNA samples using Illumina TruSeq stranded primers. RNA-libraries concentrations were measured in a Qubit fluorometer and using the Agilent 2100 Bioanalyzer.