Vertis Biotechnologie Illumina library of YALI2_RNA_TSS_POOL: 2x 75nt paired-end run
Total RNA was isolated using the RNeasy Mini Kit (QIAGEN). cDNA synthesis was performed by Vertis Biotechnologie AG. Therefore, poly(A)+ RNA was isolated from total RNA, 5' CAP species were enriched, and 5' ends were converted to 5'P ends. Afterwards, 5' Illumina sequencing adapters were ligated and the cDNA was synthesized. Finally, 200-400 bp fragments were isolated by using NEBNext dsDNA Fragmentase, 3' Illumina adapter were ligated, and PCR was carried out. Sequencing was performed using a MiSeq Reagent Kit v3 (150-cycle) in a 2x 75nt run according to the manufacturers instructions.