NextSeq 500 paired end sequencing; scRNA-seq of sheep abomasal epithelial tissue post-nematode infection
scRNA-seq of sheep abomasal epithelial tissue post-nematode infection
two sheep were infected with 50,000 Teladorsagia circumcincta infective L3 larvae. Abomasal epithelial cells were isolated separately from the two sheep using 10x Genomics single cell protocol.  Tissue sections were incubated in HBSS containing 2.5 mM EDTA, 1mM DTT and 10 g/ml DNase I, followed by HBSS containing 1U/ml Dispase II and 10 g/ml DNase I. Isolated cells were strained through a 70um Nylon mesh sterile strainer. Cell count and viability were assessed using trypan blue staining. 10x Genomics Chromium Single Cell 3' Reagent Kit (v3). Samples were subject to barcoded reverse transcription, cDNA was amplified (12 cycles) and libraries generated using 10x Genomics Chromium Single Cell 3' Reagent Kit (v3).