title: GSM7809924: Ligation; Mus musculus; RNA-Seq
library_name: GSM7809924
library_construction_protocol: Euthanized mice were perfused with PBS. Carotid arteries were extracted and placed on ice in DMEM containing 10% fetal bovine serum. To get single-cell suspensions, arteries were washed three times with PBS, cut into fine pieces, and digested for 30 minutes at 37 °C with reciprocal shaking in a mixed digesting solution containing collagenase I (1mg/ml). Single cells were resuspended in DMEM with 10% FBS. At the completion of the digestion process, all digested cells from a single sample were pooled together. Cells were collected after being filtered through a 40um cell strainer and centrifuged for 8 minutes at 500g speed at 4 °C. Subsequently, cell pellet was suspended in HBSS for further experiments. R1 (read 1 primer sequence) are added to the molecules during GEM incubation. P5, P7, a sample index, and R2 (read 2 primer sequence) are added during library construction via End Repair, A_x0002_tailing, Adaptor Ligation, and PCR. The final libraries contain the P5 and P7 primers used in Illumina bridge amplification.