GSM2574301: E14.5E_3_16; Mus musculus; RNA-Seq
After FACS sorting, single cells were manually picked up under the microscope with a mouth pipette and transferred into 4 ul of cell lysis buffer containing 0.05 ul of a 1:300,000 dilution of ERCC Spike-in RNA (Life Technologies, 4456740). The cDNA was synthesized, amplified (18 cycles) using the Smart-seq2 protocol, and purified using VAHTS DNA Clean Beads XP beads (Vazyme, N411-03). Two nanograms of cDNA were used to prepare the sequencing libraries using the TruePrep DNA Library Prep Kit (Vazyme, TD502) with 0.4 × of the standard reaction volume. PCR amplifications were conducted in 8 cycles. Purification and size-based selection of PCR products were performed to obtain libraries with a peak size of 350 bp.