GSM2232868: microconidia; Fusarium oxysporum f. sp. niveum; miRNA-Seq
Fon hyphae were isolated by filtration through a sieve (100 mesh) and microconidia were collected first by filtration then centrifugation (5000 rpm, 10 min), after washed with sterilized distilled water and immediately frozen in liquid nitrogen, stored at -80°C. Total RNA was extracted with Axyprep™ multisource total RNA miniprep kit. sRNA fraction between 18~30 nucleotides (nt) were extracted using 15% polyacrylamide / 8 M urea denaturing gel electrophoresis (PAGE) with radioactively labeled RNA as size markers. After ligated to 5’ and 3’ adapters sequentially using truncated rhythmT4 RNA ligase (Invitrogen, Carlsbad, CA), the sRNAs were reverse converted to cDNA using Superscript III reverse transcriptase (Invitrogen, Carlsbad, CA). PCR ampliﬁcation was performed to enrich the adapter-modiﬁed cDNA libraries. RNA libraries were prepared for sequencing using standard Illumina protocols