PaperRSS文献速递|优化核酸探针的DNA芯片合成和下一代测序
近年来，寡核苷酸合成和 DNA 测序技术的发展促进了人类基因组测序项目在临床和常规研究中的应用，从而促进了人类健康和我们对人类生物学的理解。
例如外显子组测序、 CRISPR 基因组编辑和 MPRA。除了对液体活组织检查进行分析之外，样品衍生细胞系、组织成像和现场分子事件调查也提供了检索生物信息的手段，不过关于次级标签的特异性的争论也是如此。
本研究在前人工作的基础上，利用限制性内切酶和滚动环扩增(RCA)等分子生物学工具在检测分析中提高信噪比的活性，开发了一种新的探针型2倍探针，通过靶标识别和多步酶转化，产生单链环状报告分子。我们特别研究了探针的性能，在单核苷酸分辨率和库方式下，通过改变茎结构中的9-nt 序列来实现。
Recent development in oligonucleotide synthesis and DNA sequencing technologies have facilitated advancements for application of the human genome sequencing project into the clinics and routine research thus advance human health and our understanding of human biology. Examples are exome sequencing, CRISPR genome editing and MPRA. Aside from analysis of liquid biopsies, sample-derived cell lines, tissue imaging and investigation of molecular events in situ also provides a means to retrieve biological information however argument on specificity from the secondary labels. Present study built on previous work on utilizing several molecular biology tools, such as restriction enzymes and rolling circle amplification (RCA) for enhancing signal-to-noise ratio in a detection assay - a novel probe-type 2 fold probe is developed to generate a single strand circular reporter molecule upon target recognition and multi-step enzymatic conversion. We specifically investigated probe performance by varying a 9-nt sequence within the stem structure at single nucleotide resolution and in library fashion.
doi: https://doi.org/10.1101/2021.05.04.442640
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