Illumina: pooling samples and loading cartridges

In this video, you will follow Dr Paola Niola preparing a sequencing library for Illumina methodology.

Libraries prepared using technologies that attach Illumina adaptors to the fragmented DNA are sequenced on Illumina platforms. Libraries made from different samples that have had unique indexes (barcodes) incorporated are pooled together to go onto the same sequencing run. Tip: libraries can be pooled prior to a final clean-up and the pool cleaned, rather than cleaning individual libraries and then pooling. The final pool is quantified by Qubit and the peak size is determined by fragment analysis e.g. TapeStation.

The double-stranded DNA is denatured, diluted to the appropriate loading concentration and pipetted into the cartridge, which is loaded into the sequencer, along with the flow cell and buffer bottle (if applicable).

Let us know in the comments if you have had any experience of loading Illumina cartridges.