GSM8077866: A Study on COL6 in periodontitis_3-day post inflammation_Col6a2-KO replicate 1; Mus musculus; RNA-Seq
Maxillae were dissected 1 day and 3 days after ligature placement. After dissection, gingival tissue was carefully removed using a scalpel under a stereomicroscope. Subsequently, the 1st, 2nd and 3rd molars were extracted from ligature side and collected into 1X PBS in a petri dish. The teeth were washed three times in 1X PBS prior to Liberase digestion (Roche, CA, USA) that was used to liberate cells from the the PDL. Liberase powder was reconstituted with 0.25% trypsin to a final concentration of 500ug/ml. The teeth were incubated in 2 ml digestion buffer for 90 min at 37°C with shaking.  After digestion, cells were centrifuged at 1500 rpm for 5 min at 4°C. The pellet was resuspended in culture medium (10% FBS α-MEM) and passed through a 70-μm cell strainer (Fisher Scientific) and then subjected to single-cell analysis. Single-cell RNA sequencing was performed using 10X Genomics Chromium instruments.