GSM3660883: AK: 2400_REP1; Nicotiana tabacum; RNA-Seq
Leaf harvests at day 3 were done at 10 h and 15 h after dawn and at day 4 at 9 h after dawn. Frozen leaf tissue (40 - 100 mg) from three independent replicates each of bacteria-primed and mock treated plants was ground (2 x 45 s, maximum frequency) by a Retsch mill (MM 400, Retsch, Haan, Germany). Each harvest was a single young systemic leaf from a single rosette. The harvested systemic leaves were all developmentally younger than the previously locally infected leaves. RNA was extracted with the RNeasy Plant Mini Kit according to the manual (QIAGEN GmbH, Hilden, Germany). RNA was quantified with a Qubit® RNA HS Assay Kit (ThermoFisher Scientific Life Technologies GmbH, Darmstadt, Germany) and RNA quality was further assessed by agarose gel electrophoresis. RNA samples were sequenced at the Max Planck Genome Centre (MP-GC, Cologne, Germany), with an Illumina HiSeq2500 (Illumina, San Diego, CA). Single end, 100 bp reads to a minimum of 30 million (± 10%) reads per sample were undertaken using a TruSeq RNA library prepared by the MP-GC.
GEO Accession
GSM3660883