title: GSM5585767: TSS SCO5243_dN_4; Streptomyces coelicolor A3(2); RNA-Seq
library_construction_protocol: RNeasy kit (Qiagen) The triphosphates at the 5' end of 6 µg RNA samples were converted to monophosphates by using 3.75 units of RNA 5' pyrophosphohydrolase (RppH; New England BioLabs) at 30ºC for 1 hour. The 5' ends of the transcripts were ligated by the adaptor (GUUCAGAGUUCUACAGUCCGACGAUC) using 15 units of T4 RNA ligase 1 (ssRNA ligase; New England BioLabs) at 25ºC for 2 hours followed by incubation at 17ºC for 18 hours. Ligated RNA samples were purified by using RNeasy Kits and RNeasy MinElute Cleanup Kits (QIAGEN). cDNA was synthesised by using a random hexamer with an adaptor sequence (GCCTTGGCACCCGAGAATTCCANNNNNN) and SuperScript IV First-Strand Synthesis System (Invitrogen). The cDNA was amplified by using KAPA Library Amplification Kit (Roche) and index primers from TruSeq Small RNA Library Preparation Kit (Illumina). Libraries were purified by using the equal volume of AMPure XP (Beckman Coulter) twice.