GSM4261651: WT_selection_sRNA; Naumovozyma castellii; ncRNA-Seq
Total RNA was isolated from the cultures using the hot-phenol method. For small-RNA sequencing, 0.5 nM of four small RNA oligonucleotides (xtr miR-427, 5′-GAAAGUGCUUUCUGUUUUGGGCG;   dme miR-14, 5′-GGGAGCGAGACGGGGACTCACT; Synthetic_siRNA_1_Guide, 5′-UAGUGCAGGUAGGUAUUUUUGUU; Synthetic_siRNA_1_Passenger, 5′-CAAAAAUACCUACCUGCACUAUA) were added to 10 ug of total RNA for use as internal standards.  Small-RNA cDNA libraries were prepared as described (Fang and Bartel, 2015), except 9–26-nt RNAs were isolated and sequenced, using 9-nt and 26-nt RNA radiolabeled standards (5′-AAACCAGUC and 5′ -AGCGUGUACUCCGAAGAGGAUCCAAA) to follow ligations and purifications. For RNA-seq, 0.5 ng of two mRNAs (chloramphenicol and firefly luciferase) were added to 5 ug of total RNA for use as internal standards.  Ribosomal RNA was depleted from the samples using Ribo-Zero Gold rRNA Removal Kit (Yeast) (Illumina, MRZY1324).  RNA cDNA libraries were prepared using NEXTflex rapid directional mRNA-seq kit (Bioo Scientific, NOVA-5138-10).
GEO Accession
GSM4261651