高通量技术加速生物医学科学

基于新一代测序sanger测序方法也称为第一代技术。总会在平台上的是第二代测序技术。第三代测序技术包括测序核酸长链。这种技术优势以前的方法:•能够快速评估甲基化网站•设备可以便携式和低成本•但目前受到错误率很高。T下一代测序(上天)技术已经极大地增强了基因组研究的输出。桑格测序,主导该领域在上天之前,花了13年才第一个人类基因组序列。上天能做到在短短5个小时。总会在生物学和生物医学研究的概述曾经是一个现场锚定了耗时的手工任务的重量。参与和艰苦的湿实验室流程慢慢输出。但是现在,高通量技术增加了科学的速度。 In this infographic, we take a deep dive into how these technologies have changed the game. Sample/genome Fragmentation End repair & adapter ligation Clonal amplification by emulsion PCR Sequencing by ligation Pyrosequencing PPi ATP-Sulfurylase ATP Luciferase Ligase Proton detection sequencing Reversible terminator sequencing Clonal amplification by bridge PCR Cluster generation A B DNA polymerase Time Intensity DNA polymerase DNA ligase G A C A T A A C A C T A C T G T G A C A T A A C A C T A A A T T C T G T DNA polymerase H+ H+ H+ In short-read NGS, genetic material (DNA or RNA) is fragmented and oligonucleotides of known sequence are bolted on through a step called adapter ligation. This enables the fragments to interact with and be identified by the sequencing system. Sequencing by ligation doesn’t utilize DNA polymerase to create a second strand. DNA ligase’s sensitivity to basepairing mismatches is exploited instead. The fluorescence produced is used to determine the target sequence. Digital images taken after each reaction are then used for analysis. Pyrosequencing detects the generation of pyrophosphates and light release to determine whether a specific base has been incorporated in a DNA chain. Proton detection sequencing counts the hydrogen ions that are released when DNA polymerizes. pH changes are detected by semiconductorbased sensor chips and are then converted to digital readouts. Reversible terminator sequencing utilizes ‘’bridgeamplification’’. During synthesis, fragments bind to oligonucleotides on the flow cell, creating a bridge from one side of the sequence to the other, which is then amplified. The fluorescently labeled nucleotides that are added during this process are detected using direct imaging.