A Fast Solution to NGS Library Prep with Low Nanogram DNA and RNA Input

As use of NGS is becoming more widespread, more challenging samples are being used, including samples available only in small amounts. At the same time, demand is increasing for faster protocols that work reliably and do not compromise the quality of the libraries produced. 

To overcome these challenges for DNA, we have developed a fast library preparation method using novel reagents, including a new DNA polymerase that has been optimized to minimize GC bias. This method enables library construction from amounts as low as 5 ng, and is suitable for both intact and fragmented DNA.

Standard “next-generation” RNA-sequencing approaches erase RNA strand information. The polarity of the transcript is important for correct annotation of novel genes, identification of antisense transcripts with potential regulatory roles, and for correct determination of gene expression levels in the presence of antisense transcripts. To address this need we have developed a novel streamlined, low input method for Directional RNA-Sequencing that highly retains strand orientation information while maintaining even coverage of transcript expression. This highly streamlined method is based on second strand labeling and excision after adaptor ligation; allowing differential tagging of the first strand cDNA ends.