GSM8478416: Fib2 silenced N. benthamiana_replicate 3; Nicotiana benthamiana; OTHER
Total RNA was extracted from 100 mg of N. benthamiana leaf tissue and treated with on-column DNase I using the RNeasy Plant Mini Kit (Qiagen). RNA fragmentation was achieved by incubating 500-800 ng of total RNA in bicarbonate buffer (pH 9.2) at 95°C for 8 minutes, followed by ethanol precipitation for purification. Following end-repair, RNA fragments were purified using the RNeasy MinElute Cleanup Kit (Qiagen) and fragmentation was confirmed by agarose gel electrophoresis. The NEBNext Small RNA library prep (New England Biolabs) commercial protocol was used for library construction.