GB149
DNA extracts from the archaeological seeds were converted into double-stranded Illumina sequencing libraries. Samples A31-A37 libraries were built using NEBnext DNA Library Prep Mast Mix Set 2 (E6070L, New England BioLabs) with modifications described in Wales et al. (2015)3. Samples A140-A150 libraries were built using Blunt-End-Single-Tube (BEST) protocol4. Indexing of the PCR was performed on a total volume of 100/25 L: 2/0.5l of a unique index oligo (10 M) and primer IS4 (10 uM)5 for capture and shotgun sequencing respectively using AmpliTaq Gold DNA polymerase (5U/l) (Applied Biosystems). Twelve to twenty-five cycles of amplification were performed (95C for 10 minutes; 12-25x 95C for 30 seconds, 60C for 60 seconds, and 72C for 45 seconds; 72C for 5 minutes). PCR products were purified with a Qiagen QIAquick PCR Purification Kit. DNA concentration was measured using the Qubit dsDNA HS Assay Kit (InvitrogemTM) following the manufacturers protocol, and DNA was also quantified and visualized for length distribution using the High-Sensitivity D1000 DNA tapes on the TapeStation 4200 (Agilent Technologies), following the manufacturers instructions. The libraries were pooled based on index compatibility and sample molarity. Libraries for the archaeological samples were sequenced on an Illumina 2500 HiSeq platform.