RDA-seq (Next generation sequencing of Representational Difference Analysis products) of triticale leaves

RNA from triticale leaves in two varieties (HEWO, MAGNAT) in two growth phases (vegetative, generative) and grown under two cultivation conditions (drought, control) were used to Representational Difference Analysis (cDNA-RDA). Leaves under drought stress used as testers while the leaves grown in control conditions were selected as driver probes. This procedure results in accumulation of unique sequences upregulated in stress condition. The analysis was taken in three replicates for each variety (HEWO, MAGNAT) for two growth phases (vegetative, generative). The tester:driver ratio in three subsequent rounds of subtractive hybridization increased from 1:50, and 1:400 to 1:200000. Difference products obtained from the third subtraction in the size range from 200 to 800 bp were sequenced.