GSM7659074: Sp9509 small RNA 2; Spirodela polyrhiza; ncRNA-Seq
RNA was extracted from 3 biological replicates of 100mg of frozen tissue for each accession using the Quick-DNA/RNA Miniprep kit (Zymo) following manufacturer's instructions with the following modifications: frozen tissue was ground under LN2 with a mortar and pestle and resuspended in the Shield solution. It was refrozen in LN2, thawed, and treated with Proteinase K, and frozen again after the addition of lysis buffer. After thawing, samples were centrifuged at full speed, RT for 3 min. to remove debris, and the manufacturer's protocol was followed as described. Afterwards, samples were DNase treated, enriched for RNAs 17-200 nt in length, and concentrated using the RNA Clean & Concentrator kit (Zymo). Libraries were prepared with the NEXTflex Small RNA-Seq Kit v3 (Perkin Elmer) following manufacturer instructions and sequenced on a NextSeq 500 (SE 75).