title: GSM9619836: Femoral epiphyseal cells of WT mice at 12 days; Mus musculus; RNA-Seq
library_construction_protocol: The femoral epiphyseal cells were transfered in MACS Tissue Storage Solution (Miltenyi Biotec, cat. no. 130-100-008) and placed on ice. The tube was centrifuged at 50 g for 1 min at 4 °C and 2 ml preheated digestion solution ((Miltenyi Biotec, cat. no. 130-095-929), 37 °C) was added into the bowel cancer tissue（Paracancerous, lymphatic node tissue) after discarding the tissue storage buffer. Tube was incubated in a 37 °C water bath and gently pipetted three times every 5 min. Enzymatic incubation time was about 45 min. When the incubation was finished, 10% FBS (Pricella, cat. no.164210-50) was added and the solution was filtered using a 40-µm cell strainer (Corning, cat. no.CLS431750). The suspensions were centrifuged at 300 g for 3 min at 4 °C, and remove the red blood cells(Roche，cat. no. 11814389001).The suspensions were centrifuged at 300 g for 3 min at 4 °C,and the pellets were resuspended in PBS with 0.01% Bovine serum albumin (Sigma, cat. no. A1933). Then cells were loaded into microfluidic chip of Chip A Single Cell Kit v1.0 (MobiDrop (Zhejiang) Co., Ltd., cat. no. S050100301) to generate droplets with MobiNova-100(MobiDrop(Zhejiang) Co., Ltd., cat. no.A1A40001). Each cell was involved into a droplet which contained a gel bead linked with up to millions oligos (cell unique barcode). After encapsulation, droplets suffer light cut by MobiNovaSP-100(MobiDrop (Zhejiang) Co., Ltd., cat. no.A2A40001) while oligos diffuse into reaction mix. The mRNAs were captured by cell barcodes with cDNA amplification in droplets. Following reverse transcription, cDNAs with barcodes were amplified, and a library was constructed using the High Throughput Single-Cell 3' Transcriptome Kit v1.0 (MobiDrop (Zhejiang) Co., Ltd., cat. no. S050200301) and the 3' Dual Index Kit(MobiDrop (Zhejiang) Co., Ltd., cat. no. S050300301). The resulting libraries were sequenced on an Illumina NovaSeq 6000 System.