NGS library prep from ssDNA—the ssDNA-Seq Kit

Prepare high-quality libraries from single-stranded DNA (ssDNA) that are not captured during conventional double-stranded DNA (dsDNA) library preparation. The ssDNA-Seq Kit enables the construction of DNA libraries from both ssDNA and dsDNA and is ideal for samples that often contain a mixture of both DNA species, including FFPE DNA, DNA from environmental sources, and damaged or degraded DNA samples.

The kit is compatible with DNA inputs as low as 10 pg and ssDNA fragments as short as 50 bp. Unlike traditional dsDNA-seq workflows, the ssDNA-seq Kit workflow begins with the conversion of all DNA species within the sample to ssDNA through a pretreatment and heat denaturation step. Following heat denaturation, Single-Strand DNA Ligase is used to add the first adaptor to the 3’ ends of all ssDNA fragments. The library is converted to dsDNA through primer extension, and the second adaptor is added to the 5’ ends of the dsDNA fragments. Finally, Unique Dual Indexes (Cat. # 634752–634756; sold separately) are used to add the indexing sequences needed for Illumina sequencing, and the library is amplified. This simple workflow can be completed in less than 2 hr.