Peter Mouritzen
Samplix ApS, Denmark
Title: Xdropâ„¢ - Targeted Sequencing enabled into the Dark and Unknown
Biography
Biography: Peter Mouritzen
Abstract
Targeted sequencing data will never be better than the input material generated during the targeted enrichment process! While this may seem trivial, very few targeted enrichment technologies allow maintaining the integrity and quality of the DNA during enrichment. This results in both false positives and false negative results and can significantly impact conclusions. The Xdrop™ technology, a novel automated microfluidics-based targeted enrichment system, enables fast targeted enrichment while maintaining the quality of the DNA and thus makes it possible to avoid the artefacts introduced with other enrichment technologies. Here we show the Xdrop™ system being employed to sequence integrated viruses and their surrounding unknown chromosomal sequence, long GC repeats, and we show phasing of cancer mutations from sub-nanograms of DNA. Regions of 40-70 kb are enriched and sequenced using Illumina, PacBio, and Oxford Nanopore sequencing at high coverage. Apart from the Xdrop™ reagents, just 0.2-10 ng of input DNA and two adjacent 20-25 bp primers are used for the enrichment of a chromosomal region and it is therefore fast and easy to set up for a new region. The primers are located in the central part of the enriched region which means that partially unknown regions can also be enriched using the system making it relevant for regions with structural variation, CRISPR gene editing, gap closing, variable viruses or bacteria, pseudogenes etc. We also show that the Xdrop™ system can be used for general, unbiased isothermal amplification of small amounts of samples of DNA for any type of downstream sequencing.