BA Fast Highly Multiplexed Solution to NGS Library Prep with Low Nanogram DNA Input.

As the quantity of data generated per next generation sequencing (NGS) run increases and the time required per run decreases, the ability to quickly produce and track large numbers of libraries is becoming increasingly important. In addition, the ability to produce high quality libraries from limited starting material and multiple sample types, including FFPE is essential. To overcome these challenges and to minimize the bottleneck of NGS library prep, we have developed a fast, streamlined DNA library preparation method using novel reagents and adaptors. This method accommodates a wide range of sample input quantities and types including genomic, ChIP and fragmented DNA (e.g. FFPE). Data analysis of libraries constructed from as little as 250 pg of ChIP DNA show high complexity and significant overlap of target peaks with libraries made from10 ng of DNA. We have extended the utility of this library prep method by developing additional adaptor and primer reagents. These include a dual barcoding approach that is compatible with Illumina library prep and our novel NEBNext adaptor. This approach enables multiplexing of up to 96 different samples, which can be used to increase the number of samples per flow cell, and/or to identify specific samples/libraries in a lab. Together, the simple, streamlined workflow and dual barcode approach, significantly reduces the turn-around time, enabling high throughput processing of samples for clinical analysis and large scale genomics studies.