Hyb-Seq of Jacobaea analoga: adult leaf
Target genes are selected based on the transcriptome assembly of Curio rowleyanus (Senecioneae) (OneKP, code: BMSE).Baits are designed and synthesized with the parameters of 120-mer baits and 6X tiling density. Libraries are prepared using NEBNext Ultra II DNA Library Prep Kit (cat. no. E7645) and NEBNext Multiplex Oligos (cat. no. E7600), with final 14 PCR cycles. Six samples, with a total amount of 500 ng DNA, are pooled for per hybridization reaction. The hybridization is kept at 65 for 40 h, followed by final 14 PCR cycles. Sequencing is done with a paired-end 150 bp on an Illumina HiSeq 4000 platform.