title: GSM7224756: MGE_A_14pcw_942; Homo sapiens; RNA-Seq
library_construction_protocol: Human fetal tissue was collected from medical terminations of pregnancy with full donor consent, through the SWIFT human fetal tissue bank (http://www.biobankswales.org.uk/swift-research-tissue-bank), under UK Human Tissue Authority research license (No. 12457) held by Cardiff University and with ethical approval of the project from the Bro Taf local research ethics committee. Gestational age was estimated through ultrasound scan prior to the procedure in combination with measurement of fetal regions. MGE tissue was harvested from seven fetuses corresponds to approximately 10, 12-15  post conception weeks (PCW). MGEs were individually incubated at 37°C for 20 min in TrypLE Express containing Donase-α. Tissue was dissociated by trituration in 200 μl of DMEM/D to obtain a quasi-single-cell suspension. The cell concentration was calculated by manual counting and viability measured by Trypan Blue assay. Single cells were captured according to the manufacturer's protocol on a C1 Single-Cell IFC microfluidic chip designed for 10- to 17-μm cells (Fluidigm, Inc). This captured one cell in each of up to 96 capture chambers. Capture efficiency and cell viability was determined by microscopy before cDNA amplification and library preparation using the SMART-Seq 46 v4 Ultra Low Input RNA kit chemistry (Clontech) and Nextera XT DNA Library Prep Index Kit (Illumina). Single cell sequencing was carried out on HiSeq 4000 Illumina platform with the following parameters: paired-end reads, read length 2 x 75bp and read depth of approximately 1 million reads per cell.