GSM5661726: PB-01E; Mus musculus; RNA-Seq
Nuclei were isolated from a single E16.5 mouse with a hypotonic, phosphate-based, lysis buffer containing sucrose and DEPC, and fixed with a combination of DSP and methanol. Nuclei were washed and resuspended in SPBSTM buffer. ~4 million of these nuclei were processed with 2 plates of RT indexes, 2 plates of ligation indexes, and 3.5 plates of PCR indexes, as per the protocol described above. About 2000 nuclei per well were distributed into the final PCR plates. All 3.5 plates of PCR reactions were pooled and sequenced on an Illumina NovaSeq 6000 with dual index reads using the S4-200 kit and standard primers (Read1 34 cycles, Index1 10 cycles, Index2 10 cycles, Read2 100 cycles), resulting in about 7.15 billion reads in total, or 4.34 billion after removal of PCR duplicates.