NGS DNA library construction from samples with damaged DNA
The xGen ssDNA & Low-Input DNA Library Prep Kit enables library preparation from damaged samples that can be difficult to obtain sequence information. The xGen ssDNA & Low-Input DNA Library Prep Kit can create libraries in 2 hours from as little
as 10 pg of input material. Libraries can be prepared from heavily damaged samples by using IDT Adaptase technology. Unlike other library methods, the Adaptase technology generates library molecules from single-stranded DNA fragments, which allows
better recovery of sample input DNA complexity from heavily nicked and degraded samples compared to other commercially available products for Illumina® platforms. Sample types include degraded FFPE, ancient DNA, chromatin immunoprecipitation
(ChIP), and other enriched samples that have undergone DNA damage.
Rescue valuable sequencing data from low-abundance samples with ssDNA content
The xGen ssDNA & Low-Input DNA Library Prep Kit is designed for users needing to sequence samples containing ssDNA for research. These include metagenomic and viromic samples where relative abundance of both ssDNA and dsDNA phage and viral genomes
can be identified. Other compatible sample types include heat-denatured pathogenic samples, samples extracted under harsh denaturing conditions to decrease extraction bias, and oncology-related liquid biopsy samples when detection of single-stranded
cfDNA content is desired.
The workflow for this kit is shown in Figure 1. As illustrated in this diagram, samples undergo four steps:
- Adaptase technology—the Adaptase reaction simultaneously performs tailing and ligation of R2 Stubby Adapter to the 3´ ends in a highly efficient, template-independent manner.
- Extension—the extension reaction generates a second strand prior to ligation.
- Ligation—ligation reaction adds R1 Stubby Adapter to the original strand.
- Indexing PCR—PCR is used to incorporate sample indexes and sequences needed for Illumina® sequencing. Indexing primers are supplied separately.