Illumina NovaSeq 6000 paired end sequencing; RNA-Seq analysis of E15.5 mouse Insm1, Neurod1 and Pax6 knockouts vs Insm1GFP.Cre/+ controls
Embryos from Insm1GFP.Cre/+; Neurod1Lacz/+ dbl het x dbl het (OR Insm1GFP.Cre/+; Paxfl/fl dbl het x dbl het) mouse matings were collected at embryonic day E15.5, where the presence of vaginal plug at noon was considered as E0.5.  Embryos were genotyped by PCR and pancreata from Insm1GFP.Cre/+ controls  and Insm1, Neurod1, or Pax6 knockout mouse embryos were dissected out for RNA islolation. Insm1GFP.Cre/+ and Neurod1LacZ/+ mice used for experiments were on CD-1 background, and Pax6fl/fl were on a C57Black/6J background.  Insm1GFP.Cre/+ controls and Insm1-, Neurod1, and Pax6-knockout embryonic pancreata were used in the experiment. Total RNA was extracted from E15.5 mouse pancreatic tissue using Trizol LS and DNA-free RNA Kit (Zymo Research). cDNA libraries were constructed using  Illumina library prep kit at Vanderbilt Technologies for Advanced Genomics.