Improved Illumina Library prep kit is the ideal solution for the Australian Genome Research Facility

Q. What features do you appreciate in each of the new kits?

A. The new Illumina Illumina Stranded mRNA Prep, Ligation demonstrated high performance at low and high inputs with high transcript homology across the differing inputs. Further to this, the kit demonstrated high performance with lower quality RNA and a streamlined workflow which can be condensed to a single day with further processing optimisations. This shows a tremendous improvement over TruSeq™ as it would cut the processing time by 35% for 96 samples(10 hours versus 6.5 hours) using a non-optimised protocol.

The same benefits can be said for Illumina Stranded Total RNA Prep Ligation with Ribo-Zero Plus, but in addition, the universal nature of the depletion module allows all samples to be treated with the same kit and without needing stock for different depletion types. Time is saved as a single depletion is being applied rather than multiple different depletions for different species.

For Illumina DNA PCR-Free Prep, the simple nature of the protocol meant we could generate a library in as little as 1.5 – 2 hours which is a generational leap as TruSeq PCR-Free traditionally takes 1 – 1.5 days to process with mechanical shearing and additional QC checks included. The lower input workflows described in the protocol allow us to offer PCR-free to a wider range of researchers.

Overall, the Illumina protocols for all the kits we tested were well written, easy to follow and much faster than previous iterations.

Q. What are the future plans for Illumina library preparation kits at the AGRF?

A. We are planning to migrate from the existing TruSeq family to the new Illumina Prep family. This would bring several advantages to the AGRF such as;

1. Streamlined service offerings
2. Lower cost and faster preparation when compared to the TruSeq family
3. Significantly lower starting material requirements, this allows the AGRF to offer PCR-Free as a standard service for whole genome sequencing.

" The Illumina Stranded Total and mRNA preps demonstrated high performance at low and high inputs. The Illumina Ribo-Zero Plus rRNA depletion module was shown to significantly deplete rRNA content from a variety of samples. The universal nature of the depletion module allows samples to be treated with the same kit without needing stock for different depletion types. Overall the high performance, flexibility and streamlined single-day workflow make these kits an ideal solution for RNA prep."