Single_Cell RNA-Seq of Mus Musculus: adult type III taste cells
Individual type III taste cells from circumvallate taste papillae that respond to 50 mM KCl were isolated using fura-2 calcium imaging. Single cell mRNA was amplified using the aRNA method using the TargetAmp 2-Round aRNA Amplification Kit 2.0 (catalog no. TAU2R51224; Epicentre, Madison, WI). Unstranded sequencing libraries were prepared using the NEBNext mRNA LibraryPrep Master Mix Set for Illumina (catalog no. E6110; New England Biolabs, Ipswich, MA).Three libraries with indices 4, 6 and 12 were multiplexed in equmolar concentration and 100bp paired end sequencing was dome on a HiSeq 200 sequencer. Raw sequences in FastQ format were aligned to reference mouse genome (version GRCm38.p3) using the STAR aligner s using Gencode M4 as the junction annotation file.