cDNA library of senescent green leaves (Clone 29C) of P. cerasifera, replicate 2
Red and green leaves 7 and 13 weeks after emergence were collected. Overall, three biological replicates were chosen for each morph and time point and RNA isolation was performed using the SpectrumTM Plant Total RNA Kit (Sigma-Aldrich, St. Louis, MO, USA) following manufacturers instructions, but adding 2% PVP (Wt 40000, Sigma-Aldrich, St. Louis, MO, USA) to Lysis Solution. After elution DNAse I (Roche ) digestion was performed. Hence RNA was purified by phenol/chloroform extraction and ethanol precipitation following standard procedure. Twelve RNA-seq libraries were obtained using the TruSeq RNAseq Sample prep kit, according to manufacturers protocol (Illumina, San Diego, CA, USA), poly-A tail was exploited to isolate mRNA fraction from total RNA. Then mRNA was chemically fragmented and subsequently converted to cDNA. Adapters were ligated to the cDNA, and 200  25 bp fragments were gel purified and enriched by PCR. Libraries were sequenced with Illumina HiSeq2000 (Illumina).