Fast Low Input Library Preparation Method for High Coverage Whole Genome Sequencing

We present an accelerated transposase-based library prep workflow that supports whole genome sequencing from low input amounts of DNA. Sequencing libraries are prepared from 50ng genomic DNA using Agilent’s novel transposition technology (SureSelect^QXT library prep kit) and Herculase II DNA polymerase, which together provide robust yields and great coverage compared to traditional time-consuming adapter-ligation methods which typically require 1-3ug input. In less than 1.5 hours (with only 20 minutes hands-on time) and using an automation-friendly workflow, researchers can prepare dual-indexed libraries for sequencing on the Illumina HiSeq or MiSeq platforms. Compared to the competition, SureSelect^QXT libraries show more consistent fragmentation, better coverage and significantly lower AT dropout rate for genomes of low GC content such as Campylobacter jejuni (30% GC).