货号: | R300381 |
供应商: | 红荣微再 |
规格: | 48Rxns |
新一代测序(NGS)能够对许多应用进行精确和无偏差的基因组分析,包括检测染色体非整倍性,拷贝数变异(CNV)、插入和缺失。这些检测尤其适用IVF诊所的胚胎植入前基因筛查(PGS)和癌症遗传学。
所有这些应用的起始材料由于使用单细胞而受到限制,并且需要基因组的可重复性扩增作为NGS下游分析的先决条件。 PicoPLEX DNA-seq试剂盒专为满足这种不断增长的需求而开发,可帮助从单个细胞制备Illumina compatible NGS文库。
PicoPLEX™DNA-seq试剂盒使用专利技术,单管三步法3小时内完成单细胞扩增。
PicoPLEX™DNA-seq试剂盒旨在扩增单细胞基因组DNA,以便在Illumina®新一代测序(NGS)平台上可靠检测染色体非整倍体和拷贝数变异(CNV)。可应用于包括使用卵裂球和滋养外胚层细胞的植入前基因筛查(PGS)和需要单细胞扩增的循环肿瘤细胞(CTC)中的CNV检测。
PicoPLEX DNA-seq Kit
适用Illumina®NGS平台的单细胞文库制备Single Cell Library Preparation for Illumina® NGS Platforms
英文名称:PicoPLEX DNA-seq Kit
中文名称:PicoPLEX DNA-seq试剂盒
用途:单细胞DNA扩增,illumina文库制备。
成功实例:PicoPLEX DNA-seq试剂盒已成功处理单卵裂球,滋养外胚层细胞和培养细胞。
PicoPLEX DNA-seq试剂盒可为单一细胞提供稳定且可重现的DNA扩增,用于有限的测序覆盖度分析,不适用于高覆盖率的深度测序,如从头测序和/或全基因组测序。
测序平台:PicoPLEX库推荐的测序平台是Illumina MiSeq系统和HiSeq 2000/2500。要在Illumina NextSeq测序仪上使用,请在库准备之前联系技术咨询。
PicoPLEX®技术 - 曾用于微阵列(arrays)法的单细胞拷贝数变异,现在有单管形式可在Illumina NGS平台上使用。 PicoPLEX WGA(单细胞全基因组扩增)长期以来一直被全世界的IVF诊所用于植入前基因筛查和诊断,以检测染色体非整倍体和拷贝数变异。
PicoPLEX DNA-seq试剂盒简化了文库制备,整个过程在单个试管或单孔内进行,减少了错误和污染,加快了时间,并降低了成本。
PicoPLEX DNA-seq试剂盒包含48份反应,包含将48份单细胞或DNA(6 pg至60 pg)转换为illumina NGS文库所需的一切,包括双端索引(illumina indexes,放在一次性使用的微孔板中)。
Rubicon Picoplex试剂订购信息
48次反应,包含一次性使用96孔板的48个双端illumina indexes。
品牌 | 货号/新货号 | 产品描述 | 包装 |
Rubicon | RB3050/R30050 | PicoPLEX WGA Kit,array CGH、PCR分析用 | 50Rxns |
Rubicon | RB3381/R300381 | PicoPLEX DNA-seq Kit,48种双端illumina indexes | 48Rxns |
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2018年,与TAKARA继续合作,授权代理(上海)Rubicon和Cellartis 试剂。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、精准医疗、细胞治疗、器官再生四大板块的产品。
021-64091883 手机号:1500 1904 520。红荣微再-客服QQ: 2395557778 经销商专员QQ:735551118
Rubicon公司成立于2000年,总部位于美国密歇根州。产品都拥有专利技术,检控严格,拥有cGMP级生产能力,有多个OEM合作伙伴,是单细胞测序文库构建的领导品牌。2017年1月,Rubicon 成为Takara的一员,并且发展良好。
PicoPLEX DNA-seq 优点
减少ambiguity:高度可重复的CNV和非整倍体检测
减少工作流程:三步完成从单细胞到测序即用文库的制备
降低成本:单个试剂盒包含测序文库制备所需一切
减少污染和错误:文库制备在单管或单孔中完成,不需要转移
缩短结果时间:Illumina NGS文库在不到3个小时内制备好
Reduce ambiguity: highly reproducible CNV and aneuploidy detection
Reduce workflow: from a single cell to a sequencing-ready library in three steps
Reduce cost: a single kit contains everything needed to prepare a sequencing-ready library
Reduce contamination and error: library prep in a single tube or well, no transfers necessary
Reduce time to results: Illumina NGS libraries prepared in less than 3 hours
Highly Reproducible CNV Detection Over the Entire Genome:Amplified libraries from 11 individual flow-sorted H929 cells were sequenced on an Illumina MiSeq® and downsampled to 250,000 total reads. Thirty-five base single-end reads were mapped to human over the entire genome.
全基因组中高度可重复的CNV检测:11个流式分选的H929细胞扩增文库在IlluminaMiSeq®上测序获得250,000个总reads。 Thirty-five base single-end reads were mapped to human over the entire genome.
Highly Reproducible CNV Detection in Chromosome 1:In the each of the individual H929 cells (see previous slider), a 22 Mb loss and 30 Mb gain were consistently detected in chromosome 1 as indicated by the pink or blue bars.
染色体1中高度可重复的CNV检测:每个单独的H929细胞中(参见上图),如粉红色或蓝色条所示,在染色体1中一致检测到22Mb丢失和30Mb增加。
PicoPLEX DNA-seq Kit试剂盒参数
样本
上样量:该试剂盒每次反应可容纳5μL加样量(细胞和缓冲液最高2.5μL)。如果细胞在1x PBS中,则不要超过2.5μL的PBS。
适用样本:1-10个哺乳动物细胞(例如卵裂球,极体,滋养层细胞,羊膜细胞,CTC,培养细胞);分离的DNA(6 pg - 60 pg人类DNA);超过50 pg加样量是可能的,但需要在步骤6和10处优化;分选的染色体。
样本采集兼容方法:流式分选(不要固定),稀释和显微操作与PicoPLEX DNA-seq兼容。
细胞洗涤:建议在PBS(不含钙,镁和BSA)中洗涤并限制洗涤缓冲液低于2.5μL。
反应时间
PicoPLEX DNA-seq试剂盒可在<3小时内裂解细胞并准备好含indexes的NGS适用文库。
运输和存储:PicoPLEX DNA-seq试剂盒(目录号R300381)在干冰上运输。试剂盒应在抵达后保存在-20°C。
质量控制:PicoPLEX DNA-seq Kit使用新一代测序(NGS)对应规格进行测试,以确保产品质量和一致性。
安全信息:遵循标准的实验室安全程序并穿戴合适的实验服,防护眼镜和一次性手套,以确保人身安全并减少样品制备和后续扩增反应过程中潜在的交叉污染。更多信息请参阅相应MSDS。
原理和工作流程
PicoPLEX DNA-seq原理图
PicoPLEX DNA-seq基于Rubicon Genomics专利的*PicoPLEX技术,用于单细胞基因组DNA(gDNA)扩增,该技术使用多重循环的随机引物进行可重复文库构建。 PicoPLEX DNA-seq Kit遵循PicoPLEX WGA Kit的相同三步工作流程,包括细胞裂解,预扩增和最终扩增。首先,特制配方裂解细胞并打断DNA。接着,专门设计的随机引物通过对产物进行最小限度的扩增来选择性地预扩增DNA,以产生高度可重现的文库。最后,用包含Illumina双重条形码的引物进行PCR扩增得到测序就绪文库。
组份
组份 | 颜色 |
Cell Extraction Buffer细胞提取缓冲液 | 绿色 |
Extraction Enzyme Dilution Buffer 提取酶稀释缓冲液 | 紫色 |
Cell Extraction Enzyme 细胞提取酶 | 黄色 |
Pre-Amp Buffer 预扩增缓冲液 | 红色 |
Pre-Amp Enzyme 预扩增酶 | 白色 |
Amplification Buffer 扩增缓冲液 | 桔色 |
Amplification Enzyme 扩增酶 | 蓝色 |
Nuclease-Free Water 无核酸酶水 | 透明 |
Dual Index Plate 双端index 微孔板 | |
Quick Protocol 快速使用手册 |
性能
Individual Library Quantification Eliminated:
Flow-sorted unsynchronized H929 cells were amplified, pooled with constant volume, and
loaded onto a MiSeq v3 flow cell. Quantification of the libraries was unnecessary before pooling due to the highly reproducible amount of product produced by PicoPLEX DNA-seq reactions. The
columns which provided no reads were wells in which the cell was absent.
免除文库定量:
流式分选所得unsynchronized H929细胞扩增后,以恒定体积pooling,并且加载到MiSeq v3流通池上。由于PicoPLEX DNA-seq反应产生高度可重复的产物,所以pooling之前无需对文库进行定量。图中空白柱子部位表示该孔不含细胞。
PicoPLEX DNA-seq Kit Provides Aneuploidy Calls Concordant with Arrays:PicoPLEX DNA-seq Kit was tested for equivalency to 24sure™ arrays (Illumina) by using the same single embryo cell library for array and NGS testing. Libraries were pooled in groups with 48 compatible bar codes, and sequenced in a 2x100 MiSeq v2 PE experiment.
Sequencing results were concordant with array results and sequencing showed a terminal gain in 2p (shown in figure) and a terminal loss in 12q (data not shown), which were consistent with an undetected translocation in the paternal DNA confirmed by high resolution FISH analysis. (Data provided by GIVF).
PicoPLEX DNA-seq试剂盒提供的染色体非整倍体检测与arrays(阵列)分析一致:通过使用相同的单胚胎细胞文库进行阵列和NGS检测,PicoPLEX DNA-seq试剂盒与24sure™阵列(Illumina)等效。文库以48个兼容的条形码pooling,并在2x100 MiSeq v2 PE中进行测序。
测序结果与阵列结果一致,测序显示2p(如图)的末端增益和12q末端丢失(数据未显示),这与通过高分辨率FISH分析确认的亲本DNA中未检测到的易位一致。(数据由GIVF提供)。
PicoPLEX DNA-Seq Kit引用文献
• New protocol based on massive parallel sequencing for aneuploidy screening of preimplantation human embryos
• Vendrell et al. 2017 Systems Biology in Reproductive Medicine, DOI: 10.1080/19396368.2017.1312633
• Construction of thousands of single cell genome sequencing libraries using combinatorial indexing
• Vitak et al. bioRxiv Jul. 23, 2016; doi: http://dx.doi.org/10.1101/065482.
• A linkage map for the Newt Notophthalmus viridescens: Insights in vertebrate genome and chromosome evolution
• Keinath et al. 2014 Developmental Biology
Rubicon 试剂订购信息
品牌 | 货号 | 新货号 | 产品描述 | 包装 |
Rubicon | RB4406 | R400406 | ThruPLEX DNA-seq 48D Kit ThruPLEX通用文库构建试剂盒 | 48Rxns |
Rubicon | RB4407 | R400407 | ThruPLEX DNA-seq 96D Kit ThruPLEX通用文库构建试剂盒 | 96Rxns |
Rubicon | RB4427 | R400427 | ThruPLEX DNA-seq 48S Kit ThruPLEX通用文库构建试剂盒 | 48Rxns |
Rubicon | RB4428 | R400428 | ThruPLEX DNA-seq 12S (48)Kit ThruPLEX通用文库构建试剂盒 | 48Rxns |
Rubicon | RB4523 | R400523 | ThruPLEX DNA-seq 6S (12 )Kit ThruPLEX通用文库构建试剂盒 | 12Rxns |
Rubicon | RB4490 | R400490 | ThruPLEX Plasma-seq 12S Kit ThruPLEX血浆测序试剂盒 | 12Rxns |
Rubicon | RB4491 | R400491 | ThruPLEX Plasma-seq 48S Kit ThruPLEX血浆测序试剂盒 | 48Rxns |
Rubicon | RB4492 | R400492 | ThruPLEX Plasma-seq 96D Kit ThruPLEX血浆测序试剂盒 | 96Rxns |
Rubicon | RB4584 | R400584 | ThruPLEX Tag-seq 6S(12)Kit | 12Rxns |
Rubicon | RB4585 | R400585 | SMARTer® ThruPLEX® Tag-seq 48S Kit | 48Rxns |
Rubicon | RB4586 | R400586 | SMARTer® ThruPLEX® Tag-seq 96D Kit | 96Rxns |
Rubicon | RB3050 | R30050 | PicoPLEX WGA Kit,array CGH、PCR分析用 | 50Rxns |
Rubicon | RB3381 | R300381 | PicoPLEX DNA-seq Kit,48种双端illumina indexes | 48Rxns |
红荣微再(上海)生物工程技术有限公司2018年,与TAKARA继续合作,授权代理(上海)Rubicon和Cellartis 试剂。
021-64091883 手机号:1500 1904 520。红荣微再-客服QQ: 2395557778 经销商专员QQ:735551118