BioMicroCenter:Nextera

Nextera DNA sample preparation for Illumina sequencing from Epicentre technologies uses a modified transposase to simultaneously fragment and tag intact genomic DNA. A limited number of PCR steps are needed to create complete Illumina libraries.

Nextera samples prepared in parallel with SPRI-prepared Illumina libraries provided a similar quality of sequencing data. DNA from the same Colobacter DNA sample was either sonicated for SPRI-prep or provided as intact DNA for Nextera prep. The samples were multiplexed and sequenced together on an Illumina GAII 40bp single-end lane. The total genomic coverage for both Nextera and SPRI-te samples was exactly the same at 97.8% coverage of Colobacter’s GC-rich genome, although complexity was greater in the sonicated samples.

The Nextera transposase does exhibit a mild GC-insertion bias, shown by the increase in percent of the first few bases. This, however, did not affect the sequencing coverage. The Nextera-prepared sample has very consistent genome-wide coverage, while the SPRI-prepared samples have a more variable coverage density.

We have used Nextera DNA sample preparation for multiple samples and have seen high-quality data for all the samples. Below is an example of a CNV study prepared with Nextera DNA preparation.