GSM4880117: WT_Single muslce fiber [s80_17722]; Mus musculus; RNA-Seq
Whole EDL muscles were carefully dissected from 2 to 3-month-old WT (n=3) and C2-/- (n=2) mice and incubated in 0.5% collagenase (Roche, 11088807001) for 90min at 37oC. Next, EDL muscles were transferred to DMEM containing 10% horse serum, and muscle fibers were released by repeated trituration using a glass pipette under a dissection microscope. An intact single fiber was selected and washed with fresh DMEM twice, followed by a final rinse with sterile PBS. The single muscle fiber was lysed in 20uL of extraction buffer, and total RNA was isolated by column purification as directed by the manufacturer (PicoPure RNA Isolation Kit, KIT0204, Applied Biosystems). The quality of the isolated RNA was checked by the Agilent 2100 Bioanalyzer using the RNA 6000 Pico Assay.