WGS E limosum: methanol
Library PreparationLibraries were prepared according to the manufacturers protocol using Nextera DNA Flex Library Preparation Kit (Illumina # 20018705). The only alterations to the protocol as outlined was the reduction of total reaction volume for processing in 96 well plate format. Library preparation and bead clean-up was run on the Mantis Liquid Handler (Formulatrix) and Epmotion (Eppendorf # 5075000301) automated platform. These programs cover Tagment Genomic DNA" to "Amplify DNA in the protocol (Mantis- Nextera DNA Flex library prep protocol) and Clean Up Libraries in the protocol (Epmotion - Library Clean Up protocol). On completion of the library prep protocol, each library was quantified and QC was performed using the Quant-iT dsDNA HS Assay Kit (Invitrogen) and Agilent D1000 HS tapes (#5067-5582) on the TapeStation 4200 (Agilent # G2991AA) as per the manufacturers protocol.Library Pooling, QC and LoadingNextera DNA Flex libraries were pooled at equimolar amounts of 2nM per library to create asequencing pool. The library pool was quantified in triplicates using the Qubit dsDNA HS Assay Kit (Invitrogen). Library QC is performed using the Agilent D1000 HS tapes (#5067-5582) on the TapeStation 4200 (Agilent # G2991AA) as per the manufacturers protocol. The library was prepared for sequencing on the NovaSeq6000 (Illumina) using NovaSeq6000 SP kit v1.5, 2 x 150bp paired end chemistry, in the Australian Centre for Ecogenomics according to manufacturers protocol.