Agilent bioanalyser high sensitivity chip

Whole-metagenome shotgun libraries were prepared in accordance with the Nextera XT DNA Library Preparation Guide from Illumina with the exception that the tagmentation time was increased to 7 min. After indexing and clean-up of the PCR products as described in the protocol, each sample was run on an Agilent bioanalyser high sensitivity chip (Agilent) to determine the size range of the fragments obtained. The concentration of the samples was also determined at this point using a Qubit High Sensitivity Assay (Life-Sciences). Samples were then pooled equimolarly and the final concentration of the pooled library was determined by quantitative PCR using the Kapa Library Quantification kit for Illumina (Roche). The pooled library was then sequenced on the Illumina NextSeq using the 2 × 150 High Output kit according to standard Illumina sequencing protocols.

Whole-metagenome shotgun libraries were prepared in accordance with the Nextera XT DNA Library Preparation Guide from Illumina, with the exception that the tagmentation time was increased to 7 min. After indexing and clean-up of the PCR products, as described in the protocol, each sample was run on an Agilent bioanalyser high sensitivity chip (Agilent Technologies Ireland Ltd., Cork, Ireland) to determine the size range of the fragments obtained. The concentration of the samples was also determined at this point using a Qubit^TM dsDNA High Sensitivity and Broad Range assay kit (Invitrogen by Thermo Fisher Scientific, Dublin, Ireland). Samples were then pooled equimolarly, and the final concentration of the pooled library was determined by quantitative PCR using the Kapa Library Quantification kit for Illumina (Roche diagnostics Ltd., West Sussex, UK). The pooled library was then sequenced on the Illumina NextSeq 500 using the 2 × 150 High Output kit according to standard Illumina sequencing protocols.