(1396-A) Walkaway automation of NEBNext UltraShear and Enzymatic Methyl-seq (EM-seq™) on the Revvity Sciclone G3 NGSx iQ workstation

Abstract: DNA methylation is an important epigenetic regulator of gene expression and is increasingly associated with development and disease. In the human genome cytosines are methylated in the CpG context and are often clustered in CpG rich regions associated with gene regulation. When compared to the traditional bisulfite conversion method, NEBNext Enzymatic Methyl-seq (EM-seq™) minimizes the damage to DNA, producing reliable libraries that enable improved detection of 5mC and 5hmC from fewer sequencing reads. However, when mechanically sheared DNA is used as starting material for EM-seq, this is a bottleneck for processing and automation of larger sample sizes. In this study, we combined and automated the new NEBNext UltraShear module (enzymatic fragmentation) with the EM-seq kit on the Revvity Sciclone® G3 NGSx iQ workstation to provide a walkway solution for laboratories. NEBNext UltraShear allows for the starting samples to be sheared to the user’s desired size on the Sciclone G3 NGSx iQ with the on deck thermal cycler (ODTC). Prepared libraries are ready for QC and sequencing after two days of sample processing on the liquid handler, including an overnight hold of samples on the ODTC after the deamination incubation.
The automated workflow offers a high throughput solution for users to process up to 96 samples per run, includes a simple user interface for selecting sample number, desired fragmentation size, and number of PCR cycles, along with deck images instructing the user through deck setup, thus reducing the chance for human errors and overall hands-on time.
For research use only. Not for use in diagnostic procedures.