High-Resolution Mapping of Bacterial Transcriptional Responses in Human-Associated Microbiota - Supplement

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Project Summary Our NIGMS-supported project aims to develop Precision Run-On and SEQuencing (PRO-seq) to capture nascent transcripts in microbial communities. Whereas we often rely on the core to provide both fragment analysis and Sanger sequencing for library preparation, these steps add considerable time to our workflow due to long queues. Fragment analysis is required to determine whether there is sufficient amount of DNA for sequencing and confirms that the size distribution is indicative of a diverse library rather than PCR artifacts such as primer-dimers. Sanger sequencing is required for work with human gut microbiota, as these organisms are rarely grown in strictly selective media, are often slow-growing, and therefore require confirmation to ensure the strain cultures are free from contamination. Here, we request support for a ThermoFisher SeqStudio which can perform both fragment analysis and Sanger sequencing, significantly streamlining our design-build-test methods development process.