Project Summary/Abstract This application seeks funding to purchase a Qiagen QIAcube Connect System. This instrument is a cutting-edge, next generation benchtop automated DNA sample preparation system configured for microbiome workflow. It will be partnered with our existing Illumina MiSeq system (ShEEP funded, 2018) and our Eppendorf epMotion automated pipetting robot to eliminate tedious manual steps in DNA extraction, increase accuracy and reliability, and maximize throughput in our 16S rRNA gene sequencing projects. Like the MiSeq, the QIAcube will support studies of the gut microbiome by our VA-funded investigators. The QIAcube enables use of well-established QIAGEN spin-column kits currently in use in our lab (manual processing) so we can continue to use QIAGEN-based chemistry to increase efficiency. The QIAcube Connect integrates a centrifuge, heated shaker, pipetting system and a robotic gripper allowing full automation of more than 80 spin columns per run. The workstation has a built-in UV light for efficient decontamination of the worktable, critical when extracting nucleotides from samples in the same workspace. The workstation contains a tablet allowing for simplified sample preparation and provides real-time feedback on run progress. The barcode scanner on the QIAcube automatically uploads kit information to the screen, streamlining run preparation. The instrument also has a variety of pre-installed protocols for DNA isolation and purification and is based on existing approaches used in our QIAGEN reagents for sample lysis, binding, washing and elution. To give an impression of the effectiveness of this instrument, a recent 16S rRNA gene sequencing study in our lab involved 350 samples (fecal pellets). The manual processing of the samples to isolate DNA took a research associate 20 workdays to complete, working 8 hours per day. The QIAcube would reduce this preparation time (40 min per 48-sample run) to ~5-6 hr of instrument time. The QIAcube system will allow expansion of ongoing, currently funded research on the following projects of our 3 major and 2 minor users: 1) The Kuhn lab is studying the role of the gut microbiome in Gulf War Veterans Illness as well as in repetitive mild traumatic brain injury; 2) the Mateika lab is studying the role of serotonin in respiratory function in spinal cord injury and how respiratory and sleep alterations associated with spinal cord injury are modulated by the gut microbiome; 3) the Rossi lab is studying the role of high fructose intake via the diet in regulating blood pressure responses to salt, and we have already started a collaborative project with Dr. Rossi on this subject; 4) the Theis lab is collaborating with the PI on a VA funded project to study the role of the gut microbiome in Gulf War Veterans Illness. Dr. Theis is also the resident expert on 16S rRNA gene sequencing and bioinformatics and his expertise is critical to this program of research. Dr. Theis is a VA researcher in our R&...