Abstract 3542: Turbolase: A radically streamlined high throughput sample prep to sequencing workflow

Normalase is a novel enzymatic library normalization method that eliminates library quantification and manual concentration adjustment of each sample prior to library pooling, and the uniform sample processing eliminates numerous error-prone pipetting steps. When combined with Swift 2S Turbo rapid library kits, a highly streamlined ‘Turbolase’ workflow is created that is readily automated on the Hamilton Star and other platforms, where simple bulk processing improves throughput and reduces cost for NGS laboratories. Swift 2S Turbo kits comprise two enzymatic steps and a single purification that completes DNA fragmentation, end repair and adapter ligation. This is followed by standard library amplification using Normalase PCR primers to condition the libraries and produce a required minimum yield in excess of the 4 nM final concentration. This is followed by a single purification and two 15-minute Normalase incubation steps that; 1) enzymatically select 4 nM of each library, and 2) enzymatically normalize each library to 4 nM within a single pool. The pools can then be directly sequenced without further purification. Normalase allows for greater than 10-fold variation in input quantity, while generating less than 10% variation in sample representation within a pool resulting in optimal cluster density and sample balance for Illumina sequencing. Because of the unique workflow, Normalase also eliminates index hopping for libraries sequenced on patterned flow cells. The ‘Turbolase’ workflow is compatible with full-length indexed adapters that have been added by ligation as well as workflows that require indexing PCR primers to complete library construction during library amplification. Normalase is also compatible with library preparation kits available from other vendors. Normalase for pooling of libraries for pre-hybridization capture is currently under development, which will make multiplexed hybridization capture protocols streamlined and robust from library generation to sequencing. Citation Format: Dustin Masser, Robert Stedtfeld, Justin Lenhart, Vladimir Makarov, Laurie Kurihara. Turbolase: A radically streamlined high throughput sample prep to sequencing workflow [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3542.