GSM5506478: Emerged haustoria-PARE/Degradome bioreplicate 1; Cuscuta campestris; OTHER
Total RNA was isolated using TRIzol reagent (Life Technologies, Carlsbad, CA, USA). Small RNAs were gel purified in a 15% acrylamide/8M urea gel. Purified sRNAs were used as input to generate sRNA libraries using the NEBNext Small RNA Library Prep Set for Illumina (New England Biolabs, Ipswich, MA, USA) following the manufacturer instructions. This procedure is identical to the one described in Martinez et al. (2016). PARE libraries were constructed following the protocol described in Zhai et al. (2014). sRNA libraries were generated using the standard NEBNext Small RNA Library Prep Set for Illumina (New England Biolabs, Ipswich, MA, USA) protocol. PARE libraries were constructed following the protocol described in Zhai et al. (2014) which, briefly, is based on the enrichment of 5'P-mRNA molecules that are ligated to an RNA adapter. After reverse transcription the adapter is used to generate 20 nt fragments using a restriction enzyme. These fragments are then ligated again to adapters that are compatible with standard sRNA Illumina primers and indexes.