title: RNA-Seq of EGFP
design: For Next-generation sequencing, the EGFP+/mCherry+ cells were sorted and collected by FACS analysis (MoFlo, Beckman). Then, total RNA was extracted from EGFP+/mCherry+ cells with the BioZol Total RNA Extraction Reagent (Biomiga) according to the manufacturers protocol. The total RNA was reverse-transcribed into cDNA with HiScript III 1st Strand cDNA Synthesis Kit (+ gDNA wiper) (Vazyme). The target EGFP fragment was amplified via PCR using Phanta Max Master Mix (Vazyme) with the following primer pairs: EGFP_FW: 5-CACCATCGTGGAACAGTACG-3; EGFP_RV: 5-GTGCAGATGAACTTCAGGGTC-3. The PCR products were purified and analysed by Next-generation sequencing with the Novaseq PE250 Platform (GENEWIZ).