GSM2332649: 37_BS-seq_culture_rep2; Picea abies; Bisulfite-Seq
DNA was extracted from needles of the sequenced Z4006 Picea abies genotype, as described in Nystedt et al 2013 Nature. For the SE culture sample, material was collected at the proliferation stage, as described in Businge et al 2012 Tree Physiol, and DNA extracted as for the needle samples. The culture was generated from seeds obtained from the sequenced ‘Z4006’ genotype. BS-seq libraries were prepared using the TruSeq DNA LT kit (Illumina) as described previously Du J et al., (2014) Mol Cell, except that EZ DNA Methylation-Lightning Kit (Qiagen) was used for bisulfite conversion of the DNA. BS-Seq libraries were sequenced on a HiSeq 2500 system (Illumina) to obtain single-end 100bp reads per manufacture instructions. For BSPCR-seq, genomic DNA were bisulfite treated by EZ DNA Methylation-Lightning Kit (Zymo) and then amplified by locus-specific primers. Amplification products were used for library generation. Libraries in repliate 1 were generated by Ovation Ultralow V2 kit (NuGen) and libraries in replicate 2 were generated by TruSeq Nano DNA LT kit (illumina). BSPCR-seq libraries were sequenced on a HiSeq 2000 system (Illumina) to obtain single-end 100bp reads per manufacture instructions.