Ku86 抗体 (B-1) Ku86 Antibody (B-1)

Ku86 抗体 (B-1)

Ku86 Antibody (B-1)

详细描述:
The Ku protein is localized in the nucleus and is composed of subunits referred to as Ku70 (p70) and Ku86 (p86) which is also known by the synonym Ku80 or (p80). Ku was first described as an autoantigen to which antibodies were produced in a patient with scleroderma polymyositis overlap syndrome, and was later found in the sera of patients with other rheumatic diseases. Both subunits of the Ku protein have been cloned, and a number of functions have been proposed for Ku, including cell signaling, DNA replication and transcriptional activation. Ku is involved in Pol II-directed transcription by virtue of its DNA binding activity, serving as the regulatory component of the DNA-associated protein kinase that phosphorylates Pol II and transcription factor Sp. Ku proteins also activate transcription from the U1 small nuclear RNA and the human transferrin receptor gene promoters. A Ku-related protein designated the enhancer 1 binding factor (E1BF), composed of two subunits, has been identified as a positive regulator of RNA polymerase I transcription initiation.

应用范围:WB, IP, IF, IHC(P), FCM, ELISA;反应种属:human;Isotype:mouse monoclonalIgG1 κ;标记:未标记。

货号 产品名称 品牌 购买
货号 名称 单位 购买
sc-5280 Ku86 抗体 (B-1) 200 µg/ml 咨询客服

Ku-86 (B-1)抗体(琼脂糖偶联) Ku-86 (B-1) AC

Ku-86 (B-1)抗体(琼脂糖偶联)

Ku-86 (B-1) AC

详细描述:
The Ku protein is localized in the nucleus and is composed of subunits referred to as Ku-70 (p70) and Ku-86 (p86) which is also known by the synonym Ku-80 or (p80). Ku was first described as an autoantigen to which antibodies were produced in a patient wi

应用范围:WB, IP, IF, IHC(P), FCM, ELISA; 反应种属:human; Isotype:mouse monoclonal IgG1; 标记:Agarose。

货号 产品名称 品牌 购买
货号 名称 单位 购买
sc-5280 AC Ku-86 (B-1)抗体(琼脂糖偶联) 500 µg/ml, 25% ag 咨询客服

Ku-86 (B-1)抗体 (HRP 标记) Ku-86 (B-1) HRP

Ku-86 (B-1)抗体 (HRP 标记)

Ku-86 (B-1) HRP

详细描述:
The Ku protein is localized in the nucleus and is composed of subunits referred to as Ku-70 (p70) and Ku-86 (p86) which is also known by the synonym Ku-80 or (p80). Ku was first described as an autoantigen to which antibodies were produced in a patient wi

应用范围:WB, IP, IF, IHC(P), FCM, ELISA; 反应种属:human; Isotype:mouse monoclonal IgG1; 标记:HRP。

货号 产品名称 品牌 购买
货号 名称 单位 购买
sc-5280 HRP Ku-86 (B-1)抗体 (HRP 标记) 200 µg/ml 咨询客服

Ku-86 (B-1)抗体 (FITC 标记) Ku-86 (B-1) FITC

Ku-86 (B-1)抗体 (FITC 标记)

Ku-86 (B-1) FITC

详细描述:
The Ku protein is localized in the nucleus and is composed of subunits referred to as Ku-70 (p70) and Ku-86 (p86) which is also known by the synonym Ku-80 or (p80). Ku was first described as an autoantigen to which antibodies were produced in a patient wi

应用范围:WB, IP, IF, IHC(P), FCM, ELISA; 反应种属:human; Isotype:mouse monoclonal IgG1; 标记:FITC。

货号 产品名称 品牌 购买
货号 名称 单位 购买
sc-5280 FITC Ku-86 (B-1)抗体 (FITC 标记) 200 µg/ml 咨询客服

Aluminium Standard Solution (Al 1000) 铝标准溶液(Al 1000) 品牌:Wako


品牌:Wako
CAS No.:
储存条件:25℃以下
纯度:990~1010mg/l
产品编号

(生产商编号)

等级 规格 运输包装 零售价(RMB) 库存情况 参考值

016-15471

JCSS (Japan Calibration Service System) 100 ml 咨询


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* 零售价、促销产品折扣、运输费用、库存情况、产品及包装规格可能因各种原因有所变动,恕不另行通知,确切详情请联系上海金畔生物科技有限公司。

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Ku-86 (B-1)抗体 ( PE 标记) Ku-86 (B-1) PE

Ku-86 (B-1)抗体 ( PE 标记)

Ku-86 (B-1) PE

详细描述:
The Ku protein is localized in the nucleus and is composed of subunits referred to as Ku-70 (p70) and Ku-86 (p86) which is also known by the synonym Ku-80 or (p80). Ku was first described as an autoantigen to which antibodies were produced in a patient wi

应用范围:WB, IP, IF, IHC(P), FCM, ELISA; 反应种属:human; Isotype:mouse monoclonal IgG1; 标记:PE。

货号 产品名称 品牌 购买
货号 名称 单位 购买
sc-5280 PE Ku-86 (B-1)抗体 ( PE 标记) 200 µg/ml 咨询客服

Ku-86 (B-1)抗体(Alexa Fluor® 488 标记) Ku-86 (B-1) Alexa Fluor® 488

Ku-86 (B-1)抗体(Alexa Fluor® 488 标记)

Ku-86 (B-1) Alexa Fluor® 488

详细描述:
The Ku protein is localized in the nucleus and is composed of subunits referred to as Ku-70 (p70) and Ku-86 (p86) which is also known by the synonym Ku-80 or (p80). Ku was first described as an autoantigen to which antibodies were produced in a patient wi

应用范围:WB, IP, IF, IHC(P), FCM, ELISA; 反应种属:human; Isotype:mouse monoclonal IgG1; 标记:AF488 。

货号 产品名称 品牌 购买
货号 名称 单位 购买
sc-5280 AF488 Ku-86 (B-1)抗体(Alexa Fluor® 488 标记) 200 µg/ml 咨询客服

Ku-86 (B-1)抗体( Alexa Fluor® 647 标记) Ku-86 (B-1) Alexa Fluor® 647

Ku-86 (B-1)抗体( Alexa Fluor® 647 标记)

Ku-86 (B-1) Alexa Fluor® 647

详细描述:
The Ku protein is localized in the nucleus and is composed of subunits referred to as Ku-70 (p70) and Ku-86 (p86) which is also known by the synonym Ku-80 or (p80). Ku was first described as an autoantigen to which antibodies were produced in a patient wi

应用范围:WB, IP, IF, IHC(P), FCM, ELISA; 反应种属:human; Isotype:mouse monoclonal IgG1; 标记:AF647 。

货号 产品名称 品牌 购买
货号 名称 单位 购买
sc-5280 AF647 Ku-86 (B-1)抗体( Alexa Fluor® 647 标记) 200 µg/ml 咨询客服

Ku-86 (B-1)抗体( Alexa Fluor® 405 标记) Ku-86 (B-1) Alexa Fluor® 405

Ku-86 (B-1)抗体( Alexa Fluor® 405 标记)

Ku-86 (B-1) Alexa Fluor® 405

详细描述:
The Ku protein is localized in the nucleus and is composed of subunits referred to as Ku-70 (p70) and Ku-86 (p86) which is also known by the synonym Ku-80 or (p80). Ku was first described as an autoantigen to which antibodies were produced in a patient wi

应用范围:WB, IP, IF, IHC(P), FCM, ELISA; 反应种属:human; Isotype:mouse monoclonal IgG1; 标记:AF405 。

货号 产品名称 品牌 购买
货号 名称 单位 购买
sc-5280 AF405 Ku-86 (B-1)抗体( Alexa Fluor® 405 标记) 100 µg/2 ml 咨询客服

SMART-Seq HT Kit & SMART-Seq HT PLUS Kit酶试剂盒Takara

上海金畔生物科技有限公司代理Takara酶试剂盒全线产品,欢迎访问官网了解更多产品信息。

SMART-Seq HT Kit & SMART-Seq HT PLUS Kit
品牌 Code No. 产品名称 包装量 价格(元) 说明书 数量
Clontech R400748 SMART-Seq® HT PLUS Kit 48 Rxns ¥26,309 SMART-Seq HT Kit & SMART-Seq HT PLUS Kit SMART-Seq HT Kit & SMART-Seq HT PLUS Kit SMART-Seq HT Kit & SMART-Seq HT PLUS Kit
Clontech R400749 SMART-Seq® HT PLUS Kit 96 Rxns ¥51,365 SMART-Seq HT Kit & SMART-Seq HT PLUS Kit SMART-Seq HT Kit & SMART-Seq HT PLUS Kit SMART-Seq HT Kit & SMART-Seq HT PLUS Kit
Clontech R400744 Unique Dual Index Kit – 48U 48 Rxns 询价 SMART-Seq HT Kit & SMART-Seq HT PLUS Kit SMART-Seq HT Kit & SMART-Seq HT PLUS Kit SMART-Seq HT Kit & SMART-Seq HT PLUS Kit
Clontech R400745 Unique Dual Index Kit – 96U 96 Rxns 询价 SMART-Seq HT Kit & SMART-Seq HT PLUS Kit SMART-Seq HT Kit & SMART-Seq HT PLUS Kit SMART-Seq HT Kit & SMART-Seq HT PLUS Kit
Clontech 634455 SMART-Seq® HT Kit 12 Rxns ¥4,458 SMART-Seq HT Kit & SMART-Seq HT PLUS Kit SMART-Seq HT Kit & SMART-Seq HT PLUS Kit SMART-Seq HT Kit & SMART-Seq HT PLUS Kit
Clontech 634456 SMART-Seq® HT Kit 48 Rxns ¥16,049 SMART-Seq HT Kit & SMART-Seq HT PLUS Kit SMART-Seq HT Kit & SMART-Seq HT PLUS Kit SMART-Seq HT Kit & SMART-Seq HT PLUS Kit
Clontech 634436 SMART-Seq® HT Kit 480 Rxns ¥118,883 SMART-Seq HT Kit & SMART-Seq HT PLUS Kit SMART-Seq HT Kit & SMART-Seq HT PLUS Kit SMART-Seq HT Kit & SMART-Seq HT PLUS Kit
Clontech 634437 SMART-Seq® HT Kit 96 Rxns ¥30,910 SMART-Seq HT Kit & SMART-Seq HT PLUS Kit SMART-Seq HT Kit & SMART-Seq HT PLUS Kit SMART-Seq HT Kit & SMART-Seq HT PLUS Kit
Clontech 634438 SMART-Seq® HT Kit 192 Rxns ¥57,064 SMART-Seq HT Kit & SMART-Seq HT PLUS Kit SMART-Seq HT Kit & SMART-Seq HT PLUS Kit SMART-Seq HT Kit & SMART-Seq HT PLUS Kit
Clontech 634439 SMART-Seq® HT Lysis Components 96 Rxns ¥1,199 SMART-Seq HT Kit & SMART-Seq HT PLUS Kit SMART-Seq HT Kit & SMART-Seq HT PLUS Kit SMART-Seq HT Kit & SMART-Seq HT PLUS Kit
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SMART-Seq HT Kit & SMART-Seq HT PLUS Kit 延伸     SMART-Seq HT Kit & SMART-Seq HT PLUS Kit SMART-Seq HT Kit & SMART-Seq HT PLUS Kit
SMART-Seq HT Kit & SMART-Seq HT PLUS Kit SMART-Seq HT Kit & SMART-Seq HT PLUS Kit
         
SMART-Seq HT Kit & SMART-Seq HT PLUS Kit
 
 
SMART-Seq HT Kit & SMART-Seq HT Plus Kit:自动化友好型的单细胞mRNA全长分析
· 自动化友好型 - 兼容自动化工作站,更少的时间分析更多的样本
· 高灵敏度 - 可以从单细胞或10 pg总RNA起始(起始量:1-100个细胞或10 pg-1 ng总RNA)
· 高质量的RNA-Seq数据 - 保留全长基因信息,低rRNA比率,高基因检出数,有效扩增GC-rich转录本
· 单管操作流程 - SMART-Seq HT PLUS Kit包含完整的cDNA合成和文库构建流程,分别在单管中进行,减少样品损失与混淆,降低手动操作误差
· 增强的测序性能 - SMART-Seq HT PLUS Kit整合双端特殊index(UDI),即便多个文库pool后用高通量型illumina测序仪(如Novaseq)分析,也能得到正确结果
· 更高的文库产量 - 更便于使用高通量型illumina测序仪测序(如Novaseq),或保存剩余的文库用于多次测序
 
产品说明
SMART-Seq HT Kit(SS-HT)和SMART-Seq HT Plus Kit(SS-HT PLUS)均兼容自动化移液工作站,采用oligo(dT)引物,支持1-100个细胞或10 pg-1 ng总RNA起始。
SS-HT试剂盒是从第4代的SMARTer Ultra Low试剂(SMART-Seq v4)发展而来的,具有简化的操作流程,将原有的反转录和PCR扩增步骤整合为一步,且不影响性能。细胞分离或RNA提取后,仅通过一步RT-PCR扩增双链cDNA,然后进行纯化完成操作。cDNA合成可在4小时内完成,整个文库构建方案可在2天内完成。而且,SMART-Seq HT试剂盒具有与SMART-Seq v4试剂盒相同的灵敏度和可重复性。核心的SMART技术,是一种对全转录组研究,以及包括细胞内剪接位点,可变剪接研究的非常强大的工具,且可以检测全长基因。 Oligo(dT)可以从高品质总RNA(RIN> 8)或完整细胞中特异性扩增mRNA。 通过采用LNA技术,提升了模板转换的效率与基因检出数等性能。
SS-HT PLUS Kit在包含核心的cDNA合成试剂基础上,同时整合了Takara Bio Group专利性的文库构建技术(U.S. Patents 7,803,550, 8,399,199, 8,728,737, 9,598,727
,10,196,686, 10,208,337, and 11,072,823及单次使用的双端特殊index(UDI),提供完整的单细胞cDNA合成到文库构建的实验体验。建库试剂中包含片段化酶,以实现对合成cDNA的片段化,接着利用特别的茎环形接头构建高品质illumina文库。在建库阶段,单管两步操作、过程中无需纯化、2 h即可建成文库!
 
SS-HT Kit:流程缩短、性能不变!
SMART-Seq HT Kit & SMART-Seq HT PLUS Kit
图1 SMART-Seq HT的技术流程
 
SMART-Seq HT Kit & SMART-Seq HT PLUS Kit
图2 SMART-Seq v4和SMART-Seq HT工作流程的比较
 
SMART-Seq HT(右)是由SMART-Seq v4方法(左)经过改进而来的一个手动操作时间更短的高通量工作流程。使用FACS方法获得单细胞,SMART-Seq HT试剂盒只需3个操作步骤即可制备cDNA,而SMART-Seq v4试剂盒需要6步。SMART-Seq HT工作流程的一个重要步骤是一步RT-PCR,即使用一步RT-PCR缓冲液,进行高效的反转录和PCR扩增cDNA的过程。
 
SMART-Seq HT Kit & SMART-Seq HT PLUS Kit
图3 使用SMART-seq v4 kit和SMART-Seq HT kit从FACS分选的293T细胞中获得的高重复性的基因表达数据
 
使用SMART-Seq v4试剂盒(SSv4_1-SSv4_12)和SMART-Seq HT试剂盒( HT_1-HT_9)(上图)从21个293T细胞单细胞制备文库,并对每个细胞获得的基因表达性进行评估。层次聚类热图展示了所有细胞之间的Euclidean distances,并显示从0.74到0.97的Pearson相关性。不仅用相同试剂盒制备的细胞之间可以观察到非常高的相关性,而在两种试剂盒之间的相关性也非常高。这些数据表明,SMART-Seq HT的工作流程与SMART-Seq v4相比,在基因表达分析中没有显著的偏差。
 
SS-HT PLUS Kit:不只是一个Kit,更是完整的单细胞RNA-Seq建库体验!
为了使单细胞cDNA合成、文库构建实验“一步”到位,我们全新推出了SS-HT PLUS Kit。
SMART-Seq HT Kit & SMART-Seq HT PLUS Kit
 
SMART-Seq HT Kit & SMART-Seq HT PLUS Kit
 
从高灵敏度、高重复性的cDNA合成出发,构建更高品质illumina文库
 
Takara科学家从10 pg小鼠脑total RNA起始,采用SMART-Seq HT Kit完成cDNA合成。随后分别使用SS-HT Plus Kit中的SMART-Seq Library Prep Kit模块与市面上较常用的A公司试剂盒构建文库,并比较性能差异。
 
SMART-Seq HT Kit & SMART-Seq HT PLUS Kit
(比较数据来源于Takara Bio USA, Inc.)
 
从测试数据中可以看出,SS-HT PLUS Kit所建文库的产量是A公司Kit产量的5倍,而更高的文库产量更适于高通量型Illumina测序仪,如Novaseq。
 
两种方法所建文库随后用Nextseq 500完成测序,并对TPM(Transcripts per million)≥0.1的基因数进行统计。SS-HT PLUS文库能比A公司文库多检出10%的基因,从同样的cDNA起始能“捕获”更全的信息!
 
 
产品详情请点击:SMART-Seq HT Kit & SMART-Seq HT PLUS Kit
 
更多实验数据:
SS-HT Kit
SS-HT Plus Kit
 
延伸阅读:
单细胞NGS解决方案
 
 

页面更新:2022-09-02 10:46:54

Skp1 p19 (H-6)抗体 Skp1 p19 (H-6)

Skp1 p19 (H-6)抗体

Skp1 p19 (H-6)

详细描述:
The critical role that the family of regulatory proteins known as cyclins plays in eukaryotic cell cycle regulation is well established. The best characterized cyclin complex is the mitotic cyclin B/Cdc2 p34 kinase, the active component of MPF (maturation

应用范围:WB, IP, IF, IHC(P), ELISA; 反应种属:m, r, h; Isotype:mouse monoclonal IgG2b; 标记:无标记。

货号 产品名称 品牌 购买
货号 名称 单位 购买
sc-5281 Skp1 p19 (H-6)抗体 200 µg/ml 咨询客服

Skp1 p19 (H-6)抗体(琼脂糖偶联) Skp1 p19 (H-6) AC

Skp1 p19 (H-6)抗体(琼脂糖偶联)

Skp1 p19 (H-6) AC

详细描述:
The critical role that the family of regulatory proteins known as cyclins plays in eukaryotic cell cycle regulation is well established. The best characterized cyclin complex is the mitotic cyclin B/Cdc2 p34 kinase, the active component of MPF (maturation

应用范围:WB, IP, IF, IHC(P), ELISA; 反应种属:m, r, h; Isotype:mouse monoclonal IgG2b; 标记:Agarose。

货号 产品名称 品牌 购买
货号 名称 单位 购买
sc-5281 AC Skp1 p19 (H-6)抗体(琼脂糖偶联) 500 µg/ml, 25% ag 咨询客服

Skp1 p19 (H-6)抗体 (FITC 标记) Skp1 p19 (H-6) FITC

Skp1 p19 (H-6)抗体 (FITC 标记)

Skp1 p19 (H-6) FITC

详细描述:
The critical role that the family of regulatory proteins known as cyclins plays in eukaryotic cell cycle regulation is well established. The best characterized cyclin complex is the mitotic cyclin B/Cdc2 p34 kinase, the active component of MPF (maturation

应用范围:WB, IP, IF, IHC(P), ELISA; 反应种属:m, r, h; Isotype:mouse monoclonal IgG2b; 标记:FITC。

货号 产品名称 品牌 购买
货号 名称 单位 购买
sc-5281 FITC Skp1 p19 (H-6)抗体 (FITC 标记) 200 µg/ml 咨询客服

Skp1 p19 (H-6)抗体 ( TRITC 标记) Skp1 p19 (H-6) TRITC

Skp1 p19 (H-6)抗体 ( TRITC 标记)

Skp1 p19 (H-6) TRITC

详细描述:
The critical role that the family of regulatory proteins known as cyclins plays in eukaryotic cell cycle regulation is well established. The best characterized cyclin complex is the mitotic cyclin B/Cdc2 p34 kinase, the active component of MPF (maturation

应用范围:WB, IP, IF, IHC(P), ELISA; 反应种属:m, r, h; Isotype:mouse monoclonal IgG2b; 标记:TRITC 。

货号 产品名称 品牌 购买
货号 名称 单位 购买
sc-5281 TRITC Skp1 p19 (H-6)抗体 ( TRITC 标记) 200 µg/ml 咨询客服

9-Anthrylmethyl N,N-Diethylcarbamate 9-蒽甲基 N,N-二乙基氨基甲酸酯(WPBG-018) 品牌:Wako


品牌:Wako
CAS No.:
储存条件:2-10℃
纯度:
产品编号

(生产商编号)

等级 规格 运输包装 零售价(RMB) 库存情况 参考值

356-33641

1 g 1,350.00


* 干冰运输、大包装及大批量的产品需酌情添加运输费用


* 零售价、促销产品折扣、运输费用、库存情况、产品及包装规格可能因各种原因有所变动,恕不另行通知,确切详情请联系上海金畔生物科技有限公司。

产品描述相关资料下载相关产品浏览记录 请联系客服

DNA-PKCS (G-4)抗体 DNA-PKCS (G-4)

DNA-PKCS (G-4)抗体

DNA-PKCS (G-4)

详细描述:
The phosphatidylinositol kinase (PIK) family members fall into two distinct subgroups. The first subgroup contains proteins such as the PI 3- and PI 4-kinases and the second group comprises the PIK-related kinases. The PIK-related kinases include Atm, DNA

应用范围:WB, IP, IF, ELISA; 反应种属:human; Isotype:mouse monoclonal IgG1; 标记:无标记。

货号 产品名称 品牌 购买
货号 名称 单位 购买
sc-5282 DNA-PKCS (G-4)抗体 200 µg/ml 咨询客服

PLC γ2 (B-10)抗体 PLC γ2 (B-10)

PLC γ2 (B-10)抗体

PLC γ2 (B-10)

详细描述:
Phosphoinositide-specific phospholipase C (PLC) plays a critical role in the initiation of receptor mediated signal transduction through the generation of the two second messengers, inositol 1,4,5-triphosphate and diacylglycerol from phosphatidylinositol

应用范围:WB, IP, IF, IHC(P), ELISA; 反应种属:m, r, h; Isotype:mouse monoclonal IgG1; 标记:无标记。

货号 产品名称 品牌 购买
货号 名称 单位 购买
sc-5283 PLC γ2 (B-10)抗体 200 µg/ml 咨询客服

PLC γ2 (B-10)抗体(琼脂糖偶联) PLC γ2 (B-10) AC

PLC γ2 (B-10)抗体(琼脂糖偶联)

PLC γ2 (B-10) AC

详细描述:
Phosphoinositide-specific phospholipase C (PLC) plays a critical role in the initiation of receptor mediated signal transduction through the generation of the two second messengers, inositol 1,4,5-triphosphate and diacylglycerol from phosphatidylinositol

应用范围:WB, IP, IF, IHC(P), ELISA; 反应种属:m, r, h; Isotype:mouse monoclonal IgG1; 标记:Agarose。

货号 产品名称 品牌 购买
货号 名称 单位 购买
sc-5283 AC PLC γ2 (B-10)抗体(琼脂糖偶联) 500 µg/ml, 25% ag 咨询客服

PLC γ2 (B-10)抗体 (HRP 标记) PLC γ2 (B-10) HRP

PLC γ2 (B-10)抗体 (HRP 标记)

PLC γ2 (B-10) HRP

详细描述:
Phosphoinositide-specific phospholipase C (PLC) plays a critical role in the initiation of receptor mediated signal transduction through the generation of the two second messengers, inositol 1,4,5-triphosphate and diacylglycerol from phosphatidylinositol

应用范围:WB, IP, IF, IHC(P), ELISA; 反应种属:m, r, h; Isotype:mouse monoclonal IgG1; 标记:HRP。

货号 产品名称 品牌 购买
货号 名称 单位 购买
sc-5283 HRP PLC γ2 (B-10)抗体 (HRP 标记) 200 µg/ml 咨询客服

SMART-Seq Stranded Kit酶试剂盒Takara

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SMART-Seq Stranded Kit
品牌 Code No. 产品名称 包装量 价格(元) 说明书 数量
Clontech 634442 SMART-Seq® Stranded Kit 12 Rxns ¥12,631 SMART-Seq Stranded Kit SMART-Seq Stranded Kit SMART-Seq Stranded Kit
Clontech 634443 SMART-Seq® Stranded Kit 48 Rxns ¥41,939 SMART-Seq Stranded Kit SMART-Seq Stranded Kit SMART-Seq Stranded Kit
Clontech 634444 SMART-Seq® Stranded Kit 96 Rxns ¥55,549 SMART-Seq Stranded Kit SMART-Seq Stranded Kit SMART-Seq Stranded Kit
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SMART-Seq® Stranded Kit
SMART-Seq® Stranded Kit:以单细胞起始,构建高质量的链特异性RNA-seq文库
· 操作简便:以1–1,000个细胞 或10 pg-10 ng的总RNA起始,仅需7h即可完成文库制备;
· 高灵敏度:可以单细胞起始,保留链特异性,最终获得全长总RNA序列信息;
· 高再现性:准确检测包含编码和非编码RNA在内的全部转录组;
· 用途广泛:适用于降解的样本;
· 适用对象:人、小鼠、大鼠。
 
SMART-Seq® Stranded Kit能够直接从1-1,000个完整细胞(或10 pg-10 ng总RNA)起始,制备链特异性Illumina测序文库。本试剂采用随机引物法可以制备完整的转录组文库,并适用于部分降解及低品质的RNA。此外,本试剂采用专有的ZapR技术,在文库制备过程中去除核糖体cDNA。
 
如果是以oligo-dT引物起始对微量高品质的RNA进行cDNA合成时,推荐使用 SMART-Seq v4 Ultra Low Input RNA Kit for Sequencing 或SMART-Seq HT Kit试剂盒。
 
SMART-Seq Stranded Kit
图1. SMART-Seq Stranded Kit的技术流程
 
SMART-Seq Stranded Kit
图2. 使用SMART-Seq Stranded Kit制备文库的结构特征
 
接头序列包含:通过PCR 1反应添加的可以与Illumina流动池结合生成簇的P7、P5序列(P7以淡蓝色表示,P5以红色表示),用于区分单个lane上多个样本的Index序列(Index 1 [i 7] 序列连接在P7端,Index 2 [i 5]连接在P5端)。并且还包括识别测序引物区域的Read 2(紫色)和 Read 1(绿色)。通过Read 1起始得到的原始RNA的反义链序列,Read 2起始获得的是原始RNA的正义链序列。其中,通过Read 2起始时,前3个碱基(XXX)是SMART-Seq Stranded Adapter衍生的序列。因此在双端测序时,需要在mapping前去掉这3个碱基。
 
SMART-Seq Stranded Kit
表1.不同样本起始量的分析结果
 
以人脑总RNA 10 pg至10 ng不同量起始,使用SMART-Seq Stranded Kit制备RNA-Seq文库。该表中所显示的数据是三次重复的平均值,且重复之间相关性系数非常高(Pearson/Spearman表示)。即使起始量为10 pg时,也具有非常高的重现性。
 
SMART-Seq Stranded Kit
图3. 在低起始量时,SMART-Seq Stranded Kit和Pico v2的比较
 
Panel A.以人脑总RNA (50 pg-10 ng)起始,分别使用SMART- Seq Stranded Kit和SMARTer Stranded Total RNA-Seq Kit v2 -Pico Input Mammalian (Pico v2)制备RNA-Seq文库(均做三次重复)。从测序数据随机抽取250万paired-end reads进行生物学信息分析。
 
Panel B.比较这两个试剂盒在总RNA以 50 pg起始制备文库的转录表达水平 (见Panel A), SMART-Seq Stranded Kit (Pearson =0.97)的再现性高于Pico v2 (pearson =0.92)。
 
SMART-Seq Stranded Kit
 
SMART-Seq Stranded Kit
图4. 不同起始量样本的高重现性
 
利用FACS分离出A375细胞,分别以1-1,000个细胞起始使用SMART-Seq Stranded Kit制备测序文库。其中,以5-1,000个细胞起始时,均做两个重复;以单个细胞起始时,做12个重复。为了作比较,同时使用从两种1,000个细胞中纯化的总RNA制备文库。
 
Panel A.:1-1,000个细胞起始的分析结果;
 
Panel B.:通过聚类热图展示了Panel A.中所示的所有样本的Euclidea距离,以及Pearson相关系数(范围为0.85至0.99)。其中单细胞样本标记为Cell1-Cell12,其他样本分别标记为a-b。
 
TechnoteSMART-Seq Stranded Kit
 
产品详情请点击:SMART-Seq Stranded Kit
 
 

页面更新:2020-04-10 11:06:07