RNA-seq of YQ1B5
Total RNA was isolated from breast samples using TRIzol (Invitrogen, CA, USA) reagent. Next, the quantity and purity of RNA were quantified using NanoDropND-1000 (NanoDrop, Wilmington, DE, USA). The integrity of RNA was confirmed by agarose gel electrophoresis. Using Oligo (dT) (ThermoFisher, USA) magnetic beads to specifically capture mRNA containing PolyA. Then, the fragmented RNA is synthesized into CDNA through the action of InvitrogenSuperScriptmll reverse transcriptase (CA, USA). Double stranded synthesis using Escherichia coli DNA polymerase I (NEB, USA) and RNaseH (NEBUSA). Later, these composite double stranded DNA and RNA were transformed into DNA double stranded. Then, the second strand was digested using UDG enzymes (NEB, MA, US), and PCR was performed to form a library with a fragment size of 300bp+50bp. Finally, double ended sequencing was performed using Iumina novaseg 6000 (LC Biotechnology Co., Ltd., Hangzhou, China).