产品推荐     
产品推荐
 
产品推荐
   
EpiNext 高灵敏染色质免疫共沉淀测序技术(ChIP-Seq)试剂盒(Illumina)(货号:P-2030-12)
发布时间:2017-11-7 16:52:34 点击:406
      染色质免疫共沉淀及后续的测序提供了一个研究蛋白质-DNA相互作用的强有力的工具,可以检测活细胞内结合在特定序列上的蛋白。现有的ChIP-Seq方法在研究蛋白-DNA互作过程中起到了重要的作用,但也有一些缺点:1. 需要大量的细胞或组织产生足够的DNA文库,这些方法不能应用于含有有限DNA量的样本,如组织活检,胚胎组织等;2. 背景很高;3. 耗时(大于3d),不方便。为了解决这些问题,Epigentek研发了EpiNext高灵敏染色质免疫共沉淀测序(Illumina)试剂盒。

  • 高效率——阳性DNA和阴性对照产物之比大于500;
  • 高灵敏——高特异性和灵活性可以用亚纳克级别的起始DNA进行非条形码序列(单个)和条形码序列(多重)DNA文库构建,每次反应用50000个细胞即可,范围为50,000-1,000,000个细胞,选择范围大 ;
  • 操作快捷——快速的实验方案,从细胞、组织到DNA文库只要不到7h ,本试剂盒含有ChIP-Seq的全部必要组分 ;
  • 误差小—— 超级HiFi扩增使得实验者可以在最小误差的情况下得到最大的DNA文库产率。

Fig.1. Workflow of the  EpiNext™ ChIP-Seq High-Sensitivity Kit (Illumina).


Fig.2. Highly sensitive ChIP: The sheared chromatin isolated from different amounts of MBD-231 cells was used for ChIP-qPCR analysis of RNA polymerase II enrichment in GAPDH promoters.

Fig.3. Size distribution of library fragments. Ten nanograms of DNA was ChIPed by RNA polymerase II enrichment and used for DNA library preparation.

Fig.4. A ChIP-sequencing library was prepared using the EpiNext ChIP-Seq High Sensitivity Kit (Cat. #P-2030) from rat heart chromatin and polyclonal antibodies against H3K4me3 (Cat. #A-4033), H3(K9/14)ac (Cat. #A-4021) and H3K27me3 (Cat #A-4039). Sequencing was carried out on an Illumina HiSeq2500. Bioinformatics analysis of ChIP-seq is performed utilizing Bowtie and MACS. ChIP-seq reveals that the peaks of H3K4me3 and H3(K9/14)ac, which are indicators of gene activation presence in the exon CpG islands (CGI) of transcription factor gene Nr1h2 (nuclear orphan receptor member), while H3K27me3, which causes gene silencing, is not enriched in the same gene. It is reported that H3K4me3 is often enriched in CGI of orphan genes.




   
上海快博生物技术有限公司
 地址:上海市闵行区新骏环路115号2号楼406-407室
 电话:021-62968268             传真:021-62968198
 邮编:201114                      E-MAIL:kbbiosh@163.com
全国服务热线:021-62968268