Illumina TruSeq RNA文库制备试剂盒v2_化工仪器网

来自 : 发布时间：2025-06-15

mRNA信使RNA（英语：messenger RNA，缩写：mRNA），是由DNA经由转录而来，带著相应的遗传讯息，为下一步转译成蛋白质提供所需的讯息。可用来研究基因表达调控。PCR聚合酶链式反应（英文：Polymerase chain reaction，缩写PCR），是一种分子生物学技术，用于扩增特定的DNA。在基因文库构建中，可以从微量起始DNA样本中得到测序所需大量DNA产物。TruSeq RNA文库制备试剂盒v2英文名称：TruSeq RNA Library Preparation Kit v2中文名称：TruSeq RNA文库制备试剂盒v2Illumina TruSeq RNA文库制备试剂盒v2 可以从总RNA生成mRNA的测序文库，用于后续的簇生成和DNA测序。Illumina测序流程：文库构建簇生成（illumina测序平台特配试剂）上机测序TruSeq RNA Library Preparation Kit v2借助增强的多重分析功能和提供预混试剂的简单工作流程，可从总RNA生成专注于mRNA的测序文库。TruSeq RNA文库制备试剂盒v2提供24个*的标签，具备更强的多重分析性能，可处理大量样本（例如，可在HiSeq 2500系统单次运行中处理多达384个RNA-Seq样本）。与之前的方法相比，预先混合试剂消除了多数移液步骤并减少了纯化量，同时zui大限度地缩短了手动操作时间。如此，可通过NGS平台提供的zui易用样本制备工作流程实现经济实惠的高通量RNA测序研究。 Simple Workflow for RNA and DNA:流程简化Master-mixed reagents and minimal hands-on steps. Scalable and Cost-Effective Solution:方案精简Optimized formulations and plate-based processing enables large-scale studies at a lower cost. Enhanced Multiplex Performance:高通量多路运行Twenty-four adaptor-embedded indexes enable highthroughput processing and greater application flexibility. High-Throughput Gene Expression Studies:高通量基因表达研究Gel-free, automation-friendly RNA library preparation for rapid expression profiling. TruSeq RNA Library Preparation Kit v2参数Assay Time实验时间~10.5 hoursHands-On Time手动时间~4.5 hoursInput Quantity加样量0.1 - 1 ug total RNA or 10 - 400 ng previously isolated mRNA (from species with polyA tails)Content SpecificationsCaptures the coding transcriptome (without strand information)Mechanism of ActionmRNA提取方法Oligo-dT beads capture polyA tailsdT磁珠抓取含polyA尾的mRNAMultiplexing多线程处理Up to 24-plex per lanezui高24通道System Compatibility兼容系统Genome Analyzer IIx, HiSeq 1000, HiSeq 1500, HiSeq 2000, HiSeq 2500, HiSeq 3000, HiSeq 4000, MiSeq, NextSeq 500, NextSeq 550Species物种Bovine, Human, Mammalian, Mouse, Other, RatVariant ClassGene Fusions, Novel Transcripts, Single Nucleotide Polymorphisms (SNPs), Transcript VariantsTechnologySequencingMethod方法mRNA SequencingmRNA 测序Automation CapabilityLiquid Handling Robots Truseq RNA文库构建原理dT磁珠抓取含polyA尾的mRNA，从总RNA中提取mRNA。逆转录技术合成cDNA。PCR法扩增DNA产物。 TruSeq RNA Library Preparation Kit v2组份和保存Box A: 保存 -15 to -25 CBox B: 保存 -15 to -25 CBox A和 Box B主要作用：cDNA末端修复。注：根据需要选择Box A或 Box B。Box A和 Box B的区别是RNA接头Index不同。Box 1: 保存 2 to 8 C，主要成分：RNA 纯化磁珠。Box 2: 保存 -15 to -25 C 主要成分：磁珠清洗，cDNA模板合成。PCR Prep Box: 保存 -15 to -25 C。主要作用：PCR法富集产物。 TruSeq RNA文库制备试剂盒v2加样量要求TruSeq RNA Sample Preparation v2 protocols are optimized for 0.1 1 µg of total RNA.Lower amounts of RNA might result in inefficient ligation and low yield.优化加样量：总RNA0.1-1µgRNA量低于0.1µg可能会影响连接效率导致产量低。You can also use previously isolated mRNA as starting material. Use the entire fraction of mRNA purified from 0.1 1 µg of total RNA.也可以预先从总RNA分离得到mRNA作为起始样本。从0.1-1µg总RNA分离的所有mRNA都要用于加样。 Illumina® TruSeq® RNA Sample Preparation Kit v2制备流程This protocol explains how to convert the mRNA in total RNA into a library of template molecules suitable for subsequent cluster generation and DNAsequencing using the reagents provided in the Illumina® TruSeq® RNA Sample Preparation Kit v2.The first step in the workflow involves purifying the poly-Acontaining mRNA molecules using oligo-dT attached magnetic beads. Following purification, the mRNA is fragmented into small pieces using divalent cations under elevated temperature. The cleaved RNA fragments are copied into first strand cDNA using reverse transcriptase and random primers. Second strand cDNA synthesis follows, using DNA Polymerase I and RNaseH.The cDNA fragments then go through an end repair process, the addition of a single A base, and then ligation of the adapters. The products are then purified and enriched with PCR to create the final cDNA library.流程概述：使用Illumina® TruSeq® RNA Sample Preparation Kit v2试剂将总RNA的mRNA转化为模板文库，以用于后续的DNA测序。1.纯化：通过oligo-dT磁珠法纯化得到poly-A mRNA分子。2.mRNA 切段：高温，二价阳离子。3.mRNA 段子转为*条cDNA：逆转录酶，随机引物。4.第二条cDNA合成：DNA 聚合酶I ，RNaseH。5.cDNA段子末端修复：添加A碱基，连接接头。6.产物纯化，PCR富集，得到cDNA文库。TruSeq® RNA Sample Preparation Kit v2文库制备方法分为低样品量和高样品量两种。 TruSeq® RNA Sample Preparation Kit v2特点Multiplexing capability with 12 indexing adapters for a total of 24 adapters Ability to process up to 384 samples per run on a HiSeq 2000Optimized fill volumes Automation support Streamlined workflow:Master-mixed reagents reduce reagent containers and pipettingUniversal adapter for preparation of mRNA samplesHigh throughput:Enables simultaneous preparation of multiplexed mRNA samplesVolumes are optimized for standard 96-well plateTroubleshooting with built-in process control checksUniversal index adapter tags for all samples. Additional adapters and primers are not necessary. TruSeq RNA和DNA文库制备相关产品RNA文库制备TruSeq RNA Library Preparation Kit v2, Set A(12 indexes, 48 samples) RS-122-2001TruSeq RNA Library Preparation Kit v2, Set B(12 indexes, 48 samples) RS-122-2002DNA文库制备TruSeq DNA Library Preparation Kit v2, Set A(12 indexes, 48 samples) FC-121-2001TruSeq DNA Library Preparation Kit v2, Set B(12 indexes, 48 samples) FC-121-2002For Cluster Generation on cBot and Sequencing on the HiSeq2000/1000 and HiScanSQTruSeq Paired-End Cluster Kit v3 cBot HS(1 flow cell) PE-401-3001TruSeq Single-Read Cluster Kit v3 cBot HS(1 flow cell) GD-401-3001For Cluster Generation on cBot and Sequencing on the Genome AnalyzerIIxTruSeq Paired-End Cluster Kit v2 cBot GA(1 flow cell) PE-300-2001TruSeq Single-Read Cluster Kit v2 cBot GA(1 flow cell) GD-300-2001For Cluster Generation on the Cluster Station and Sequencing on the Genome AnalyzerIIxTruSeq Paired-End Cluster Kit v5 CS GA(1 flow cell) PE-203-5001TruSeq Single-Read Cluster Kit v5 CS GA(1 flow cell) GD-203-5001mRNA测序简介mRNA测序(mRNA-Seq)已迅速成为分析疾病状况、生物过程及广泛研究设计中的转录组的方法。mRNA-Seq不仅可提供极为准确且高灵敏度的量化基因表达，还可识别已知的和新的转录异构体、基因融合和其他特征及等位基因特异性表达。mRNA-Seq可提供编码转录组的完整视图，而并不受限于先验知识。mRNA测序的优势与基因表达芯片相比，mRNA-Seq在分析转录组方面具有诸多优势：该方法适用于更广泛的动态范围，不仅可增加灵敏度，还可提高基因表达中测量的倍数变化的准确性。可捕获已知特征和新发特征可广泛应用于众多物种用于癌症研究的mRNA-Seq癌症会积累大量的基因变化，但通常只有少数发展为肿瘤。借助mRNA-Seq监控基因表达变化有助于识别疾病预后或治疗反应的预测性生物标志物。mRNA-Seq还支持对基因表达和病因进行特征分析，并可了解影响肿瘤分类和发展的变异。用于复杂疾病研究的基因表达图谱基因表达图谱和调控研究可让您形象地了解基因组、表观遗传和环境因素对常见疾病的影响程度。mRNA-Seq是一种成熟的探查和图谱分析工具，可提供具有较高影响力的研究结果，并识别针对一系列复杂疾病的下游调查的目标靶点，其中包括神经系统、免疫系统、新陈代谢以及心血管方面的疾病。品牌货号产品描述IlluminaRS-122-2001TruSeq RNA Library Preparation Kit v2TruSeq RNA文库制备试剂盒v2，套件A（48个样本，12个标签）IlluminaRS-122-2002TruSeq RNA Library Preparation Kit v2TruSeq RNA文库制备试剂盒v2，套件B（48个样本，12个标签）TruSeq RNA文库制备试剂盒v2及更多illumina NGS测序产品欢迎您咨询华雅再生医学旗舰公司：红荣微再（上海）生物工程技术有限公司 ：1500 1904 520。红荣微再-客服:2395557778 经销商专员红荣微再以 传递科学价值，服务科学研究 为宗旨，主营干细胞、医疗、细胞治疗、器官再生四大板块的产品。 关注本网官方微信 随时阅读专业资讯