title: GSM6674875: transcription start sites from KARS-T promoter; Homo sapiens; OTHER
library_construction_protocol: Total RNA was extracted directly with Trizol. The Renilla luciferase RNA was reverse transcribed with a gene-specific primer, followed by alkaline RNA hydrolysis. The 3' end of the cDNA was then ligated to a double stranded DNA linker with a 3' NNNNNN or GNNNNN (mixed in 1:4 ratio). A 3' adapter sequence was added by PCR, followed by a second PCR that incorporates Illumina sequences with dual index. Sequencing libraries from plasmid were prepared by adding adapter sequences in a first PCR step, followed by incorporation of illumina sequences in a second PCR reaction.