Improved workflows for high throughput library preparation using the transposome-based Nextera system.

ABSTRACT:

Background

The Nextera protocol, which utilises a transposome based approach to create libraries for Illumina sequencing, requires pure DNA template, an accurate assessment of input concentration and a column clean-up that limits its applicability for high-throughput sample preparation. We addressed the identified limitations to develop a robust workflow that supports both rapid and high-throughput projects also reducing reagent costs.

Results

We show that an initial bead-based normalisation step can remove the need for quantification and improves sample purity. A 75% cost reduction was achieved with a low-volume modified protocol which was tested over genomes with different GC content to demonstrate its robustness. Finally we developed a custom set of index tags and primers

SUBMITTER: Lamble S 

PROVIDER: S-EPMC4222894 | biostudies-literature | 2013 Nov

REPOSITORIES: biostudies-literature