GSM9323685: HCT116-H2b-GFP, Stretched (+Peristalsis), Top Channel (Non-Invaded); Homo sapiens; RNA-Seq
Cells were removed from the OOC using 0.25% trypsin for 5 min at 37C added to both the top (non-invaded) and bottom (invaded) channels, repeated three times to ensure full collection.  After collection, the GFP labeled tumor cells were seclected using the FACS Melody Cell Sorter (Beckton Dickenson, Brea, CA, USA) Single cell RNA libraries were be generated using the Chromium Single Cell 3' kit (10X Genomics, CA, USA). Viability was checked with Thermo Fisher Countess II FL with Nuc blue for all cell nucleus staining and PI for dead cell nucleus staining. The PCR enzyme used was part of the 10x kit and the number of PCR cycles were 12. Loading was performed to target capture of ~10,000 GEMs per sample for downstream analysis, and samples were processed through the Chromium Controller following the standard manufacturer's specifications.  The sequencing libraries were evaluated for quality on the Agilent TapeStation (Agilent Technologies, Palo Alto, CA, USA), and quantified by using Qubit 2.0 Fluorometer (Invitrogen, Carlsbad, CA) and pooled libraries were quantified using qPCR (Applied Biosystems, Carlsbad, CA, USA) prior to loading onto an Illumina HiSeq4000 or equivalent. The samples were sequenced at a configuration compatible with the recommended guidelines as outlined by 10X Genomics.
GEO
GSM9323685_r1