title: GSM8680677: Hippo_1; Mus musculus; RNA-Seq
library_construction_protocol: The brains were dissected on ice, the cerebral cortex and hippocampus were isolated, quickly frozen using liquid nitrogen and stored at -80°C until use. Crushed tissue was added to Lysis Buffer to dissolve and extract the nuclei; Debris was removed using Debris Removal Solution (Miltenyi Biotec) and the nuclei were fixed with 80% methanol. Libraries were prepared using a single-cell combinatorial indexing RNA sequencing technique. Reverse transcription reactions were performed using 384 different barcoded Oligo dT primers to add the 1st barcode to cDNA obtained from nuclear RNA. The DNA was then distributed into 96-well plates by Pool & Sort and subjected to double-strand synthesis and Tagmentation. PCR was performed in each well using different Index primers and a 2nd barcode was added. The resulting product was column purified and small fragments were removed with AMPure XP beads.