WGBS T8 isolate, replicate 2
DNA from 9 biological replicates from culture of two culture ages (1, 8 months) and from 9 replicates of T8P were randomly selected for sequencing. Biological repli-cates were used to generate 3 pooled samples per culture age. Bisulphite treatment was performed independently from 50ng of genomic DNA of each pooled sample using the EZ DNA methylation-Gold Kit (Zymo Research) according to the manu-facturers instructions but adjusting the final column purification elution volume to 10 l. Following Bisulphite treatment, recovered DNA from each pool was used to esti-mate yield using a NanoDrop 100 spectrophotometer using the RNA setting. Bisulphite treated samples were then used to create a sequencing library using the EpiGnome Methyl-Seq Kit and Index PCR Primers (4-12) according to manufactur-ers instructions. In order to provide a reference draft sequence for the alignment of the bisulphite treated DNA and to detect any culture induced genetic mutations, 10 ng of native (non-bisulphite treated) DNA extracted from cultures from time points 1 month (T1) and 8 month (T8) were sequenced and compared to the reference B. cinerea B05.10 genome sequence. Libraries were prepared using the EpiGnome Methyl-Seq Kit and Index PCR Primers (Epicentre) (1-2) according to manufacturers instructions.