High molecular weight DNA purification instrument for metagenomics

在自然界观察到的所有微生物中，只有不到1%可以在实验室中培养，留下 研究人员无法研究某些环境中99%以上的微生物-微生物 有时具有独特的能力，如合成可用作新药或 抗生素。元基因组学，即不可培养微生物的基因组重建，是一种强大的新技术。 获取这些生物未开发资源的工具。在一种方法中，大碎片OFDMA 从含有不可培养微生物的样本中提取并克隆到宿主中，如E.coli. 来制作一个元基因组学文库。然后对文库进行筛选，以确定表达化合物的实用性。 虽然用这种方法发现了新的抗生素和酶，但成功地生产出了 由原始微生物合成的化合物，如抗生素，是一个艰巨的挑战。 克隆足够长的片段，以保持由通常超过100kb的基因簇编码的路径 长度。现有的纯化技术倾向于将基因组DNA剪切成50kb或更小的片段。这个 发现一种从土壤等困难样品中提取和纯化高相对分子质量DMA的方法将提供一种 元基因组学的突破，可能会发现许多未来的药物和抗生素。 我们最近演示了一种用于DMA浓缩的非线性电泳法，即 能够在不需要机械处理的情况下进行DMA操作和浓缩(例如离心法或 移液器)。从细菌裂解液中提取DNA，浓缩倍数达到4000倍 演示了。这种方法是下一代DNA提取方法的极佳候选者。 方法从土壤或体液等复杂样品中提取。我们建议开发一种携带 验证了该浓缩法对土壤中高分子量DNA的提取效果 旨在发现新药和抗生素的宏基因组学研究样本。对此进行许可 技术将被用于解决其他引人注目的应用，包括从身体中提取DNA 用于癌症生物标志物和病原体检测的液体。

Less than 1% of all microorganisms observed in nature can be cultured in the laboratory, leaving researchers unable to study more than 99% of microorganisms in some environments - microorganisms that sometimes have unique abilities such as synthesis of compounds that could find use as new drugs or antibiotics. Metagenomics, the genomic reconstruction of unculturable microorganisms , is a powerful new tool for accessing the untapped resources of these organisms. In one approach, large fragments ofDMA from a sample containing unculturable microorganisms are extracted and cloned into a host such as E. coli to produce a metagenomics library. The library is then screened for utility of expressed compounds. Though new antibiotics and enzymes have been discovered with this method, successful production of compounds, such as antibiotics, synthesized by the original microorganism presents the difficult challenge of cloning fragments long enough to hold pathways encoded by gene clusters that are often over 100kb in length. Existing purification techniques tend to shear genomic DMAto fragments of 50kb or less. The discovery of a method to extract and purify high MW DMAfrom difficult samples such as soil will provide a breakthrough for metagenomics that may enable the discovery of many future drugs and antibiotics. We have recently demonstrated a non-linear electrophoretic method for DMA concentration that is capable of DMA manipulation and concentration without mechanical handling (such as centrifugation or pipetting). Extraction of DNA from bacterial lysate and 4,000 fold concentration factors have been demonstrated. This method is an excellent candidate for the next generation of methods for DNA extraction methods from complex samples such as soil or body fluids. We propose to develop an instrument to carry out this concentration method, validating its performance on extraction of high molecular weight DNA on soil samples for metagenomics studies aimed at discovery of new drugs and antibiotics. Licensing of this technology will be pursued to address other compelling applications including DNA extraction from body fluids for cancer biomarker and pathogen detection.