GSM5350073: Type2_Intron1 [13707X5]; Homo sapiens; RNA-Seq
Frozen muscle biopsies (~ 4 milligrams) were cut on a cryostat (20 microns) and homogenized with a 26-gauge needle in 400 microliters of 20 mM Tris*Cl pH 7.4, 150 mM NaCl, 5 mM MgCl2, 1 mM DTT, and 1% Triton X-100.  RNA was extracted using TRIzol® and precipitated with isopropanol according to the manufacturer's instruction (Life Technologies). Total RNA (~1 microgram) was depleted of human cytoplasmic and mitochondrial rRNA using tiling oligodeoxynucleotides and the RNase H digestion method (Adiconis et al., 2013; DOI: 10.1038/nmeth.2483).  Illumina TruSeq Stranded Total RNA library kits were used to prepare indexed libraries.