GSM4946253: MuSCs, Post-injury Day 7, Sestrin1,2 Knockout [d7_SestrinKO_Rep1]; Mus musculus; RNA-Seq
Hind limb muscles (tibialis anterior, gastrocnemius, and quadriceps) of wild type and Sestrin1,2 knockout mice were dissected using sterile surgical tools at 0, 7, and 21 days after injury. Tissues were minced and transferred into 50 mL conical tubes containing 20 mL of digest solution. Cell sorting was done using a BD FACSAria III Cell Sorter. MuSCs were FACS sorted directly into Trizol and snap frozen. Samples were subsequently thawed, and RNA was extracted using a Qiagen miRNeasy Micro Kit as per manufacturer's instructions. The integrity of the isolated RNA was verified with a Bioanalyzer (Agilent 2100) and 1-10 ng of high-quality RNA (RIN>8) was used to produce cDNA libraries using the SmartSeq v4 protocol (Clontech) as per the manufacturer's instructions. cDNAs were prepared into sequencing libraries using 150 pg of full-length cDNA amplicons (Nextera XT DNA Library Preparation Kit, Illumina) with dual index barcodes. Barcoded cDNA libraries were pooled into a single tube and sequenced on a NextSeq (Illumina) using 76-bp single-ended reads.
GEO Accession
GSM4946253