GSM1202207: ppe degradome flower; Prunus persica; RNA-Seq
degradome cDNA libraries construction protocol: German MA, Luo S, Schroth G, Meyers BC, Green PJ (2009) Construction of Parallel Analysis of RNA Ends (PARE) libraries for the study of cleaved miRNA targets and the RNA degradome. Nat Protoc 4: 356-362. The construction of degradome libraries differed considerably from past efforts and followed the procedure of as Ma et al. (Ma Z, Coruh C, Axtell MJ (2010) Arabidopsis lyrata small RNAs: transient MIRNA and small interfering RNA loci within the Arabidopsis genus. Plant Cell 22: 1090-1103.) with some modifications. (1) Approximately 150 ng of poly (A)+ RNA was used as input RNA and for annealing with biotinylated random primers; (2) Strapavidin capture of RNA fragments was performed via Biotinylated Random Primers; (3) 5´ adaptor ligation was only performed to those RNAs containing 5´-monophosphates; followed by (4) reverse transcription and PCR; (5) libraries were sequenced using the 5´ adapter only, resulting in the sequencing of the first 36 nucleotides of the inserts that represented the 5´ ends of the original RNAs.