Target-enrichment sequencing of AAV-transduced frozen mouse liver
On the day of treatment, mice were weighed and given a single bolus via intravenous injection of the tail vein with the respective treatment using an injection volume of 4 L/g of body weight. For tissue collection, mice were anesthetized and exsanguinated. The median liver lobe was fixed and processed for histologic analysis. The remaining liver was split into 2 samples, snap-frozen using liquid nitrogen, and stored at 80C until all the samples were collected and processed for molecular or biochemical analysis. For each animal, 1 liver fragment, approximately 10 to 20 mg in weight, was homogenized using a stainless steel bead and 500 L RLT buffer (Qiagen, Hilden, Germany) using a TissueLyser II instrument (Qiagen). The genomic DNA and total RNA were then extracted from the same homogenate using the DNA/RNA AllPrep kit (Qiagen) following the manufacturers instructions.