GSM7978312: ENS, Plate 2 3; Mus musculus; RNA-Seq
We harvested ~30 mm of the VGLUT2-tdTomato mouse distal and gently dissected out the longitudinal and muscular layers containing the myenteric plexus, enzymatically dissociate into individual cells, and culture overnight. Under a fluorescent microscope, each tdTomato-positive neuron will be manually pipetted into a single tube containing 5 µL lysis mix. cDNA library was generated using a library preparation kit (PWSCR96, SeqWell, MA). Briefly, In the first barcoding step, a unique i7-barcoded adapter is inserted into each of the 96 cDNA samples in segregated reactions. Next, the i7-barcoded cDNA samples are pooled into a single tube. In the second barcoding step, a single i5-barcoded adapter is inserted into each pool of cDNA samples. Finally, each 96-plex library is amplified in a single PCR reaction using universal library primers.