GSM3398924: Male GMP mRNA; Mus musculus; RNA-Seq
Bone marrow was extracted by crushing hindlimbs isolated from mice immediately following euthanasia. The single cell suspension was enriched using CD117 Microbeads on an AutoMACS Pro (Mitenyi) and stained with TotalSeq antibodies and fluorchome-conjugated antibodies for FACS-sorting, then 10,000 cells per sample were loaded in each lane of a Chromium Controller Instrument (10x Genomics). The 10x Genomics single cell 3' v2 assay protocol was followed as described until the cDNA amplification step. The cDNA amplification mix was modified to include 1 mL of a 2 mM dilution of ADT PCR additive primer (5'-CCTTGGCACCCGAGAATT*C*C-3' where * indicates a phosphorotioate bond), replacing 1 mL of water. An 0.6X SPRI selection was performed, and the resulting supernatant was set aside (contains ADTs) while the beads (mRNA-derived cDNA) were processed according to the standard 10x Genomics single cell 3' v2 assay protocol. The final pooled library was comprised of 90% mRNA-derived library and 10% ADT library prior to sequencing.
GEO Accession
GSM3398924