GSM7832825: MAGL-3; Rattus norvegicus; RNA-Seq
Newly collected fresh ileal tissue were washed in ice-cold RPMI1640，cut into small pieces (<1 mm) with the sterile scissors on ice. The tissues were dissociated using Collagenase Ⅱ (Sigma, V900892-100MG), Trypsin (Hyclone, SH30042.01) and DNase I (Sigma, DN25-1G). The obtained single-cell solution was sieved through a 70 μM cell strainer (Corning). Cell count and viability was estimated using fluorescence Cell Analyzer (Countstar® Rigel S2) with AO/PI reagent after removal erythrocytes (Solarbio, R1010) Single-cell cDNA libraries were generated as described in detail (Single cell 3' whole transcriptome amplification protocol, BD Rhapsody) using successfully prepared single-cell suspensions. In brief, freshly single-cell suspensions were washed three times, and counted and subsequently loaded onto a cartridge, lysed inside the wells followed by mRNA capture on cell capture beads according to manufacturer instructions. Cell capture beads were retrieved and washed prior to performing reverse transcription and treatment with Exonuclease I. cDNA underwent extension using the WTA extension primers, purified, and then purified using SPRIselect magnetic beads (Beckman Coulter). Quantity of PCR products was determined by Qubit 4.0 with Qubit dsDNA HS Assay (Q32854)