Transcriptome changes in leaves of the DEFENSE NO DEATH 1 (DND1) silenced potato plants
Transcriptome changes in leaves of the DEFENSE NO DEATH 1 (DND1) silenced potato plants
Leaf samples for total RNA extraction were taken from a sample set of 15 leaves harvested from five 4-week-old in vitro grown plants. Plants were propagated in vitro in MS medium (doi.org/10.1111/j.1399-3054.1962.tb08052.x) without vitamins containing 2% (w/v) sucrose and 0.8% agar) at 24 °C with a 16-h photoperiod at a light intensity of 75 μmol m-2 s-1 for four weeks in 40-mL glass test tubes closed with paper plugs. Total RNA was extracted from potato leaves according to Stiekema et al. (doi: 10.1007/BF00027383). RNA concentration was measured at OD260 using an ND-1000 nanodrop spectrophotometer (Thermo Fisher Scientific, Waltham, MA, USA). Integrity of RNA was tested by the Novogene Company Ltd. (Cambridge, UK) on a gel. It was carried out by the Novogene Company Ltd. (Cambridge, UK). Messenger RNA was purified from total RNA using poly-T oligo-attached magnetic beads. After fragmentation, the first strand cDNA was synthesized using random hexamer primers, followed by the second strand cDNA synthesis using dUTP for directional library. The library was checked with Qubit and real-time PCR for quantification and bioanalyzer for size distribution detection.