Ampl-Seq of cDNA for target genes in post-culture KMST6
Amplicons prepared by RT-PCR from pre- or post-treatment sample were pooled in equimolar amounts by these sizes and digested with dsDNA Fragmentase kit to 400-500bp size. Fragmented DNA was used for library preparation using NEBNext Ultra II DNA Library Prep Kit (#E7645; NEB) with unique-dual index adaptor from NEBNext Multiplex Oligos for Illumina Kit (#E7395; NEB). Library concentration and quality were examined by DNA ScreenTape (5067-5365; Agilent) using 2200 TapeStation (Agilent). Paired-end sequencing (2  250 cycles or 2  150 cycles) was performed on Illumina Miseq with Reagent kit v2 (#MS-103-1002 or MS-103-1003; Illumina) at Tsukuba i-Laboratory LLP.