Festival of Genomics 2022 Presentation: Long-Read Sequencing Workflows with Joanna Greenhough

Here, Joanna Greenhough introduces you to some of Promega's products, which support the NGS workflow, including long-read sequencing. The process of next-generation sequencing starts with the extraction and purification of DNA samples. The next step is the amplification of genes of interest, in the process of PCR target genes, from samples are then pulled into a sequencing library. The next step, is loading the sequencing instrument. Promega's Maxwell RSC instrument is a compact automated nucleic acid purification platform, which processes up to 16 samples simultaneously, using pre-filled cartridges and pre-programmed methods. The Maxwell RSC offers consistent reliable DNA or RNA extraction in 25 to 60 minutes, depending on sample type. The process is compatible with multiple sample types such as blood, buccal swabs, cells and tissues, plants and food, fecal??? samples, FFPE saliva and plasma. Our customers refer to Maxwell as a workhorse of their lab, which enables high-quality nucleic acid extraction and purification, with minimal steps and less hands-on time and improves the quality of the extracted material. The high quality of the DNA, extracted by the Maxwell instrument, allows for the amplification of even the longest genes that are being studied. The presence of a proofreading enzyme to repair DNA mismatches and a highly processive polymerase allows the polymerase to continue to elongate the DNA much further, resulting in longer DNA amplification. High-performing polymerase is especially important when amplifying large, variable-length or hyperpolymorphic and complex genes. The next stage in the NGS process is the library preparation. ProNex size-selective purification system can be used multiple times throughout the library preparation process, from cleaning up the initial PCR reaction through to the other enzymatic reactions throughout the library prep. The ProNex system enables users to select the desired size of purified DNA fragments from 100 to 700 base pairs. Also ProNex beads have significantly lower viscosity than other beads, making them far easier to pipette, increasing reproducibility of size selection and recovery. ProNex beads can be used in both manual and automated high throughput workflows. Another Promega product that can be used in the NGS workflow is the Quantus Fluorometer, a compact easy-to-use dual-channel fluorometer designed to provide highly sensitive fluorescent detection when quantifying nucleic acids. The Quantas Fluorometer can be used at a number of stages in the NGS workflow. This is just a very short summary of the role of Promega's products in supporting the long-read sequencing workflow. Please contact Promega if you would like any additional information or have any questions about our products.