GSM3463317: Wild-type replicate 2; Porphyromonas gingivalis ATCC 33277; RNA-Seq
RNA was purified from P. gingivalis cultures that were treated with Trizol (Invitrogen) to facilitate cell lysis. Briefly, a fully grown 3 ml culture was spun and the pellet brought up in 900 μl Trizol, mixed with 200 μl chloroform and the sample spun for 15 minutes. The upper aqueous layer was transferred to a new tube, an equal volume of isopropanol added, mixed well and frozen in a -80C freezer. The next day, the samples were thawed, spun, pellet washed washed with 70% ethanol, and re-suspended in water. 20 μg of this sample, containing crude RNA, was subjected to DNase treatment using RQ1 DNase (Promega). The sample was again subjected to Trizol and chloroform treatment, spun and the upper aqueous layer was concentrated using Zymo Clean and Concentrator kit (Zymo Research), according to manufacturer's instructions. Illumina ScriptSeq kit with RiboZero