NextSeq 500 sequencing; A conserved population of MHC II-restricted, innate-like, commensal-reactive T cells in the gut of humans and mice: Human Data
A conserved population of MHC II-restricted, innate-like, commensal-reactive T cells in the gut of humans and mice: Human Data
LPMCs were isolated by DTT and EDTA treatment, digestion in a buffer containing collagenase and DNAse and a 2 layer percoll gradient. Cells  were stained with fluorecently labelled antibodies and sorted as CD161hiCD56-, CD161hiCD56+, CD161int or CD161- directly into PCR tubes containing 4µl SmartSeq2 lysis buffer. Sorted samples were spun down and cooled immediately and stored at -80 degree Celsius until RNA extraction. Reverse transcription was done directly from the sorted, lysed cells following the protocol published by Picelli et al in 2014. (https://www.nature.com/articles/nprot.2014.006) SmartSeq2 following manufacturer's instructions based on the protocol published by Picelli et al., 2014 (https://www.nature.com/articles/nprot.2014.006) following the specific SOPs of the MRC WIMM Sequencing Facility.