新一代测序
新一代测序(NGS)泛指可针对生物体指定区域或全基因组进行大规模平行或深度覆盖测序的多种相关技术。作为基于基因组学研究领域不可或缺的手段，测序技术已问世数十年之久。但直到NGS测序技术或DNA和RNA大规模平行测序技术的持续进步，研究人员才得以拓展全基因组测序覆盖度和数据分析能力，同时急剧降低测序成本。NGS应用已不再局限于全基因组分析，对于基础基因组学和疾病研究的最新进步更是影响深远。
相关技术文章
- Regardless of next-generation sequencing (NGS) platform, universal and index adapter sequences are required for the proper assembly of sample fragments.
- MISSION siRNA Universal Negative Controls are an essential component to any siRNA experiment. Using a Negative siRNA control allows the researcher to create a baseline for mRNA knockdown efficiency. MISSION siRNA Universal Negative Controls have been tested in human, rat, and mouse cells. All have been carefully designed to have no homology to known gene sequences.
- MISSION Positive Control siRNA, designed using the Rosetta siRNA Design Algorithm, serve as ideal complement to any siRNA silencing experiment. All MISSION Positive Control siRNA have been validated to work, so that you can spend more time silencing your gene of interest and less time on optimizing internal controls.
- Onxy Quencher emits heat instead of light and offers and improved signal-to-noise ratio over fluorescent quenchers.
- Next-generation sequencing (NGS) revolutionized genomic research, and is now playing a crucial role in the clinical environment.
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相关实验方案
- The SeqPlex DNA Amplification Kit for whole genome amplification (WGA) is designed to facilitate next-generation sequencing (NGS) from extremely small quantities or from degraded/highly fragmented DNA
- Annealing is the process of heating and cooling two single-stranded oligonucleotides with complementary sequences.
- The SeqPlex RNA Amplification kit provides a method for amplification of total RNA or isolated mRNA prior to entry into the workflows of the commonly used deep sequencing platforms.
- GenomePlex® is a Whole Genome Amplification (WGA) method that allows the researcher to generate a representative amplification of genomic DNA
- WTA2, a Whole Transcriptome Amplification (WTA) method, allows for representative amplification of nanogram quantities of total RNA in less than 4 hours without 3-bias
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NGS方法概览
随着NGS方法和试剂不断发展，目前涌现出众多可供研究人员选用的NGS系统。现有常用平台整合了NGS流程中的多个关键步骤，包括样品或文库制备、簇生成、测序和数据分析。样品制备通常涉及DNA扩增或添加序列接头。DNA序列簇生成指DNA共价连接接头后，与固体表面杂交进行桥式PCR扩增，或者进行微乳液PCR。现有多种DNA测序方法，包括连接法测序、合成法测序、焦磷酸测序和离子半导体测序。每种测序方法都采用不同的反应步骤和化学法，最终决定序列长度（读长）、错误率和试剂成本。
NGS数据分析方法
测序后的数据分析是所有NGS工作流程最后的关键步骤。由于每个NGS平台和工作流程都会生成海量数据信息，生物信息学家必须借助种类繁多的分析工具（如Bowtie、Galaxy等）来分析这些原始数据，进行序列比对和定位。如此庞大数据集需要通过融合各个学科来开发和优化各类分析和解读工具，这也是推动NGS技术领域不断发展的重要动力。根据特定的应用需求，这些强大的工具可帮助研究人员测序整个基因组、外显子组或转录组来进行基础和疾病研究。
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