SOPs

Genome assembly protocol

Library preparation was done using the TruSeq RNA Sample Preparation Kit v2 (Illumina). In brief, starting from total RNA the mRNA is purified using polyA selection. Following chemically fragmentation the mRNA is converted into single-stranded cDNA using random hexamer priming. Next, double-stranded cDNA is generated and adapters are ligated.

Total RNA was extracted using TriZol.

The basal 5 millimeter of the fourth leaf was sampled for RNA extraction harvested two days after its appearance from the surrounding older leaves.

Plants were grown in the growth chamber under controlled growth conditions (24oC, 55% relative humidity, 170 mmol m-2 s-1 photosynthetically active radiation at plant level in a 16-h/8-h day/night cycle).

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