WGBS of Arabidopsis Thaliana: Ler
Bisulfite-converted, Illumina sequencing libraries were constructed using reagents provided in three kits by New England Biolabs: 1) EpiMark Bisulfite Conversion Kit (E3318); 2) NEBNext Multiplex Oligos for Illumina (Methylated Adaptor, Index Primers Set 1, (Cat # E7535S); 3)NEBNext Ultra II DNA Library Prep Kit for Illumina (Cat No. E7645S). Samples (10-200 ng) were spiked-in with 1pg pUC19 (methylated control) and 20 pg Lambda DNA (unmethylated control) previous to library construction. Indexing (for multiplex sequencing of library pools) was done by 18 cycles of amplification on the cleaned, desulphonated reaction using Illumina-specific indexing primers and NEBNext Q5 Master mix. This procedure resulted in 0.5-1.0 g of library with average size of ~400 bp.