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

人类相关微生物群中细菌转录反应的高分辨率图谱 - 补充材料

• 批准号: 10825052
• 负责人: Ilana Lauren Brito
• 金额: $ 9.29万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-26 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10825052
• 关键词: Colorectal Cancer; Communities; Complex; DNA sequencing; Development; Dideoxy Chain Termination DNA Sequencing; Disease; Ensure; Genetic Transcription; Human; Human Microbiome; Inflammatory Bowel Diseases; Libraries; Maps; Metagenomics; Methods; Morphologic artifacts; National Institute of General Medical Sciences; Organism; Oxygen; Preparation; Process; Prokaryotic Cells; RNA; RNA Stability; Reaction; Resolution; Ribosomal RNA; Running; Time; Transcript; Untranslated RNA; Work; bacterial community; design,build,test; dimer; environmental stressor; gut microbiota; human microbiota; method development; microbial community; microbiome; response; transcriptome sequencing

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.

项目摘要 我们的NIGMS支持的项目旨在开发精确运行和测序（PRO-seq），以捕获 微生物群落中的新生转录物。而我们经常依赖核心来提供碎片 分析和桑格测序的文库制备，这些步骤增加了相当多的时间，我们的工作流程，由于 到排长队需要进行片段分析以确定是否有足够量的DNA用于 测序并确认大小分布指示多样性文库而不是PCR伪影 例如引物二聚体。桑格测序是研究人类肠道微生物群所必需的，因为这些微生物 很少在严格选择的培养基中生长，通常生长缓慢，因此需要确认 确保菌株培养物无污染。在此，我们请求支持ThermoFisher SeqStudio可以进行片段分析和桑格测序，大大简化了我们的工作。 设计-建造-测试方法开发过程。