Y. lipolytica producing intracellular RFP in TF Azf1 (YALI0_A16841g) knockout background (biol rep 2)
Biomass samples to be analyzed via RNAseq were rapidly centrifuged, washed once in sterile saline solution, and resuspended in TRIzol (Ambion; Thermo Fisher Scientific); then, stored at -80 C until shipment. The set of samples was forwarded to the Novogene facility for RNA isolation, quality control, library preparation, and sequencing on the Illumina platform, according to an in-house protocol. Briefly, the quality of RNA isolates was determined using BioAnalyzer (Agilent) capillary electrophoresis. mRNA was purified from total RNA using poly-T oligo-attached magnetic beads. After fragmentation, the first strand cDNA was synthesized using random hexamer primers, followed by the second strand cDNA synthesis. The library was ready after end repair, A-tailing, adapter ligation, size selection, amplification, and purification. Different libraries were pooled based on the effective concentration and targeted data amount, then subjected to Illumina sequencing.
SUB16005318
Azf1_KO_2