title: GSM4304755: 6 weeks old human placenta; Homo sapiens; RNA-Seq
library_construction_protocol: Single cells were obtained from enzymatic digestion of human first trimester placenta. Cell viability and concentration were measured by Countess II FL Automated Cell Counter (Life Technologies) targeting ≥75% cell viability. The cell emulsification is performed with the 10X Chromium Controller using the Chromium Single Cell 3' GEM Library & Gel Bead Kit v3 (10X Genomics 1000092) and Chromium Chip B Single Cell Kit (10X Genomics 1000074). The RT reaction to generate 10X barcoded single stranded cDNA, in the single cell containing GEMs, is performed using an Eppendorf MasterCycler Pro thermal cycler. Post GEM-RT Cleanup is conducted by breaking the GEMS with 10X Recovery Agent to separate the aqueous phase from the Recovery Agent and Partitioning oil. A cleanup of the sscDNA containing aqueous phase is completed using Dynabeads MyOne Silane beads (Life Technologies 37002D). The second strand cDNA amplification is performed using the cDNA Amplification Mix on the Eppendorf MasterCycler Pro. The 3' gene expression library construction is initiated with fragmentation, end repair and A-tailing of the dscDNA followed by adapter ligation and a sample index PCR using the Illumina compatible indexed adapters in the Chromium i7 Multiplex kit (10X Genomics 120262). Validation of the single cell library is conducted using the Agilent Tapestation 4200 ScreenTape assay (Agilent 5067-5576). Single Cell RNA-seq (10X Genomics Chromium Single Cell 3' v3)
attributes.tag: GEO Accession
attributes.value: GSM4304755