RubiconRB4490/R400490ThruPLEXPlasma-seq12SKit血浆游离DNA测序试剂盒基因测序试剂盒
【血浆游离DNA基因测序试剂盒主要产品】
1. ThruPLEX® Plasma-seq Kit
2. ThruPLEX® DNA-seq Kit
3. PicoPLEX® DNA-seq Kit
4. PicoPLEX® WGA Kit
5. TransPLEX® C-WTA Kit
【血浆游离DNA基▲因测序试剂盒产品应用】
1. 外显子ㄨ组富集
2. SureSelect® 富集
3. SeqCap® EZ 富集
4. 福尔马林固定石蜡包埋DNA测序
5. 胚胎植入前遗传学筛查(PGS)/种植前♂遗传学诊断(PGD)
6. DNA测序
7. RNA测序
8. 染色体免疫共沉淀测序
【产品详情】
产品名称 |
平台 |
应用程序 |
样本 |
样本量 |
技术 |
ThruPLEX®DNA-seq Kit |
Illumina NGS |
WGS, WES, 靶向测序, 染色体免疫共沉淀测序, 转录组测序 |
FFPE DNA, gDNA, cDNA, |
50 pg – 50 ng |
高灵敏度基于连接的扩增ぷ |
ThruPLEX®-FD Prep Kit |
Illumina NGS |
WGS, WES, 靶向测序,ctDNA分析 |
Cell-freeDNAisolated from plasma |
<1 ng – 30 ng |
高灵敏度基于连接的扩增 |
PicoPLEX™DNA-seq Kit |
Illumina NGS |
CNV,Aneuploidy |
Eukaryotic Cells |
<6 pg – 60pg |
准随机引物线性扩〗增 |
PicoPLEX™WGA Kit |
阵列,PCR |
CNV,Aneuploidy |
Eukaryotic Cells |
<6 pg – 60 pg DNA |
准随机引物线性扩增 |
TransPLEX®C-WTA Kit |
阵列,PCR |
基因表达分析 |
RNA |
10 – 300 ng RNA |
准随机引物扩增 |
Overview
ThruPLEX® Plasma-seq is powered by ThruPLEX chemistry to generate high performance NGS libraries from cell-free DNA isolated from plasma.
The chemistry is optimized specifically for cell-free DNA to maximize the library complexity and to preserve the GC representation of the input DNA. Libraries can be used for CNV analysis, whole genome sequencing applications or enriched using your custom panels or leading target enrichment platforms, such as Agilent SureSelect® and Roche NimbleGen SeqCap® EZ.
Libraries prepared with ThruPLEX Plasma-seq provide added sensitivity to better understand genetic variations found within the plasma samples. Plasma, along with other liquid biopsies, has gained significant interest in the research community as a valuable and easily available sample type. These communities include translational research in oncology and developmental research involving the fetal fraction derived from maternal plasma.
ThruPLEX® Plasma-seq Kit is intended for Research Use Only. It may not be used for any other purposes including, but not limited to, use in diagnostics, forensics, therapeutics, or in humans. ThruPLEX Plasma-seq may not be transferred to third parties, resold, modified for resale or used to manufacture commercial products without prior written approval of Rubicon Genomics, Inc.ThruPLEX Plasma-seq Kit is protected by U.S. Patents 7,803,550; 8,071,312; 8,399,199; 8,728,737 and corresponding foreign patents. Additional patents are pending.
Designed for cell-free DNA: newly formulated repair and ligation reagents with optimized protocols
High performance NGS libraries: high diversity with broad and reproducible GC coverage
Variable sample input: <1 ng to 30 ng of cell-free DNA
Fast and simple workflow: 3 steps in a single tube or well in 2 hours with no purification or sample transfer steps
Research applications: libraries from samples where results require high sensitivity including liquid biopsies, ctDNA, and targeted sequencing
Automation-friendly: Beckman® FXP Workstation
ThruPLEX Plasma-seq Kit generated quality libraries with high diversity and a low number of duplicates and unmapped reads. Cell-free DNA was extracted from 3 plasma samples, and libraries were prepared at the amounts indicated as measured by Qubit庐. The amount of mononucleosomal DNA in each sample as measured by the Bioanalyzer庐 corresponded to 0.09 ng, 0.62 ng, and 15.44 ng. Pooled libraries were sequenced on an Illumina NextSeq庐 500 as a paired-end run with 17M to 25M reads per library, Duplication rates were calculated after down-sampling the data to 17 M reads per library.
ThruPLEX Plasma-seq Kit libraries were captured at high efficiency and generated data with deep coverage of the kinome for mutation detection. Libraries were prepared from 3 plasma samples at input amounts of 5 ng, 6.5 ng, and 10 ng in triplicate, and targeted sequencing was carried out on an Illumina MiSeq using samples enriched with the ClearSeq Human DNA Kinome Panel for SureSelectXT2. On average, 5M reads were generated per library. Selected bases were successfully captured bases that were in or within 250 bp of the baits.
常见问题解答
1.ThruPLEX Plasma-seq可以使用什么类型的@ 样品和输入?
ThruPLEX Plasma-seq Kit旨在从分离自血浆的无细胞DNA中产生Illumina NGS的DNA文库。 ThruPLEX Plasma-seq化学■被重新制备,以产生具有广泛GC覆盖率的多种文库,提供1至30ng *无细胞DNA的可重复测序结果。
应用输入︻样品类型推荐输入量
血浆和液体活组织检查DNA测序; ctDNA分析,靶向测序从血浆中分离∑的无细胞DNA 1 ng - 30 ng *
库的输入量小于1 ng,超过30 ng。
2.ThruPLEX DNA-seq Kit和新型ThruPLEX Plasma-seq Kit之间的主要技术差异是什么?
ThruPLEX Plasma-seq Kit已经针对从血浆中分离的无细胞DNA进行△了优化。
ThruPLEX Plasma-seq Kit不需要剪切或』破碎无细胞DNA。
协议是不同的(见下文)。
3.ThruPLEX Plasma-seq和ThruPLEX DNA-seq之间的协议差异是甚么?
模板制▲备步骤:体积和保持温度
库的合成步骤:体积和培养温度
文库扩◥增步骤:扩增循环的体积和数量
4.来自ThruPLEX Plasma-seq的试剂可以与ThruPLEX DNA-seq?
不,ThruPLEX Plasma-seq Kit的内▓容物与其他Rubicon Genomics产品不可互换。
5.你有什么类型的索引或条形码?
ThruPLEX Plasma-seq 12S和48S试剂盒提供分布在管或96孔≡板中的与Illumina兼容的8nt单指数。这些索引序列由英国剑桥的Wellcome Trust Sanger研究所开∮发。关于Sanger指数序列的信息可以在Nature Methods 7,111-118(2010)中找到。
ThruPLEX Plasma-seq 96D试剂盒配备了分布在96孔板中的Illumina TruSeq HT i5和i7双重指标。 i5索引具有Illumina D501至D508序列,i7索引具有D701至D712序列。
有关索引☉的更多信息,请参阅“ThruPLEX Plasma-seq套件使◢用手册”中的附录1。
6.您有什么建议将血浆中的无细胞DNA分离出来?
使用QIAamp®循环核酸试剂盒(Qiagen,CAT.NO.55114)制备的无细胞DNA优化ThruPLEX Plasma-seq。
含有输入DNA的缓冲液中Tris和EDTA的浓度分别不得超过10mM和0.1mM。避免含磷酸盐缓冲液∩。
7.我♂应该使用什么样的收集管来吸取受试者的血液?
Rubicon Genomics的科学人员使用K2 EDTA(Becton Dickinson,CAT。NO。36643)在Vacutainer Venous Blood Collection Tubes中收集的血液∏。
8.在分离无细胞DNA之前,我应该如何在血液中制备血浆?
使用双离心方案将血▽浆与血液分离。将处理过的血浆样品在-80°处储存,直到不分离DNA。
9.从ThruPLEX Plasma-seq Kit中可以使用从尿液或其他生物流体中分离的无细胞DNA吗?
ThruPLEX Plasma-seq用从人血浆中分离的无细胞DNA进行了优化ζ ;然而,该试剂盒可能与来自其他生物流体的降解的DNA一起使用。必须←首先用DNA提取/纯化试剂盒提取其他样品来源的无细胞DNA。由于来自这些来源的无细●胞DNA通常被降解/分裂,因此不需要额外的碎裂。每个反应的无细胞输入DNA的体积不得超过10μL。
10.ThruPLEX Plasma-seq Kit是否与任何目标浓缩系统兼容?
是的,ThruPLEX Plasma-seq Kit与主要的exome和目标富集产品兼容,包括AgilentSureSelect®,RocheNimbleGen®SeqCap®EZ和定制☆面板。