GSM7846544: Sample 18_CC_FLOX_2; Mus musculus; RNA-Seq
Whole embryos were dissected from pregnant dams in 1X phoshate buffered saline (PBS). Parietal and visceral yolk sacs were removed for DNA extraction and PCR genotyping. Extraembryonic tissue was removed for genotyping by X-gal stain. Intraembryonic tissues from embryos were transferred into 1.5 ml DNA LoBind tubes, washed with 1X PBS, incubated in 1X TrypLE Express at 37°C for 20 min (cardiac crescent) min, dissociated into single cells by passage through pipette, quenched with 1X PBS, underlayed with hanks balanced salt solution + 1% deaceytlated bovine serum albumin, and spun at 1200 rpm for 5 min in a swing bucket centrifuge. The resulting upper supernatant layer was discarded by pipette and the cells were spun again at 1200 rpm for 5 min. The resulting supernatant was again discarded. The cells were washed in 1X PBS by raising them in 1X PBS, spinning them at 1200 rpm for 5 min, and discarding the resulting supernatant.  The cells were then re-suspended in 1X PBS. Single cell RNA libraries were constructed using 10X Genomics Chromium Single Cell 3' Reagents Kit v2 protocol and paired-end sequenced with single-indexing on the Illumina HiSeq 4000 to a minimum of 50,000 reads per cell using its standard protocol.