GSM9753936: Sample 594, 6 months post ASCT; Homo sapiens; OTHER
Cells are isolated from up to six 25um sections cut from rehydrated FFPE blocks using the xylene and GentleMACS OctoDissociator section of the Sample Preparation from FFPE Tissue Sections for GEM-X Flex Gene Expression protocol. Cell yield and quality is checked using NucSpot 470 stain on a Revvity Cellometer Ascend automated cell counter. If cells will not be barcoded immediately, they are stored in Long Term Storage Buffer at -80C. Fixed cells are first hybridized with Probe Barcodes. Using a microfluidic chip, the hybridized samples are then partitioned into nanoliter-scale Gel Beads-in-emulsion (GEMs). A pool of ~737,000 10x GEM Barcodes (also referred to as 10x Barcodes) is sampled separately to index the contents of each partition. Up to 16 samples may be equally pooled in each GEM reaction using uniquely barcoded probe sets for generation of one library. Final libraries are checked for size on the Agilent Tapestation and concentration using the Invitrogen Qubit 2.0. Libraries are then sequenced on a NovaSeq X Plus sequencer.