GSM6502534: Fltp+ early; Mus musculus; RNA-Seq
Islets of Langerhans have been isolated using a standard protocol (Corbin et al., 2021; https://doi.org/10.1186/s12575-021-00143-x; D.-S. Li et al., 2009; https://doi.org/10.1038/nprot.2009.150). We sorted Fltp negative (tomato positive), Fltp lineage positive (GFP positive), and Fltp transient (double positive) populations and generated separate libraries for each population. Male and female mice were pulled together after sorting. Libraries of single cells were produced using the Chromium Single-Cell 3′ library and 10XGenomics gel bead kit v3.1 (PN #1000121). Briefly, 10,000 cells were loaded per channel of a 10x chip to produce gel bead-in-emulsions (GEMs). Then the samples underwent reverse transcription to barcode RNA followed by cleanup, cDNA amplification, enzymatic fragmentation, 5′ adaptor, and sample index attachment. The samples were sequenced using a NovaSeq6000 (Illumina) with 100 bp paired-end sequencing.