手机验证
大量
Methylation Specific Bisulfite Seq Library Prep Kit
详询
北京百奥创新科技有限公司
详询
具体价格请联系我们
产品简介:
亚硫酸氢盐测序(Bisulfite seq)是一种众所周知的DNA甲基化检测技术,WGBS、RRBS、MeDIP-Seq和MSBS等技术被用于全基因组DNA甲基化分析。DNA甲基化在分子生物学、遗传学和表观遗传学中对细胞发育、分化和基因表达的调控具有重要意义。大多数甲基化胞嘧啶位于CpG位点,70-80%的胞嘧啶发生甲基化。人类基因组中CpG位点的数量约为2800万个,不到基因组的1%,而预期为4.4%。
Bisulfite seq is a well know technology to detect DNA methylation and several technologies such as WGBS, RRBS, MeDIP-Seq, and MSBS are used for whole genome DNA methylation analysis. DNA methylation is important for regulation of cell development, differentiation and gene expression in molecular biology, genetics and epigenetics. Most methylated cytosines are found at CpG sites, and 70-80% of cytosines are methylated. The number of CpG sites in human genome is around 28 million, which is less than 1% of the genome compared with 4.4% expected.
全基因组亚硫酸氢盐测序(WGBS)是目前最有效的DNA甲基化分析方法。唯一的限制是测序成本非常高,因为全基因组测序包括所有非甲基化区域。
还原重亚硫酸盐测序(RRBS)是甲基化CpG位点的一小部分还原。RRBS结合了限制性内切酶消化和亚硫酸氢盐测序,丰富了甲基化CpG位点的测序。它是一种在单个碱基水平上评估全基因组甲基化模式的有效技术。虽然这允许了更高的覆盖深度并降低了测序成本,但限制是只有10%的甲基化CpG位点被覆盖。
甲基化DNA免疫沉淀测序(Methylated DNA Immunoprecipitation Sequencing, MeDIP-Seq)是另一种用于选择甲基化DNA的全基因组富集技术。利用抗5-甲基胞嘧啶抗体,通过免疫沉淀从全基因组DNA中富集甲基化DNA。5-甲基胞嘧啶抗体与基因组片段DNA孵育并沉淀,随后进行DNA纯化和测序。MeDIP-Seq有几个缺点:1.相对于单基分辨率,低分辨率(150~200 bp);2. 由于抗体特异性和选择性的非特异性相互作用。3.偏向于高甲基化区域。
BioDynami开发了BioDynami Methylation Specific Bisulfite Seq建库试剂盒(illumina平台),以亚硫酸氢盐处理的DNA (20 ng - 500 ng)为输入,构建甲基化CpG位点的NGS文库。试剂盒富集了甲基化的CpG区域,从而显著降低了测序成本。该试剂盒基于亚硫酸氢盐测序技术,在单个碱基水平上估算全基因组甲基化模式。
亚硫酸氢盐处理完成的NGS库会对库造成巨大的破坏。通过使用亚硫酸氢盐处理过的DNA作为输入,该试剂盒克服了由于亚硫酸氢盐转化而造成的重大文库损失。该试剂盒包含PCR聚合酶的混合物,具有高保真扩增和尿嘧啶耐受性,是亚硫酸氢盐处理DNA的理想选择。
Whole genome bisulfite sequencing (WGBS) is the most effective method of DNA methylation analysis. The only limitation is the sequencing cost is very high because the whole genome is sequenced including all the non-methylated regions.
Reduced Representation Bisulfite Sequencing (RRBS) is the reduced representation of a smaller fraction of the methylated CpG sites. RRBS combines restriction enzyme digestion and bisulfite sequencing, and enriches the sequencing for methylated CpG sites. It is an efficient technology for estimate the whole genome methylation patterns at the single base level. Although this allows a higher coverage depth and reduces the sequencing cost, the limitation is only 10% of the methylated CpG sites are covered.
Methylated DNA Immunoprecipitation Sequencing (MeDIP-Seq) is another whole genome enrichment technique used for selection of methylated DNA. Using antibodies against 5-methylcytosine, methylated DNA is enriched from whole genomic DNA via immunoprecipitation. 5-methylcytosine antibodies are incubated with fragmented genomic DNA and precipitated, followed by DNA purification and sequencing. There are several drawbacks of MeDIP-Seq: 1. Low resolution (150~200 bp) as opposed to the single base resolution; 2. Non-specific interaction due to antibody specificity and selectivity. 3. Bias towards hypermethylated regions.
The Methylation Specific Bisulfite Seq (MSBS) Library Prep Kit (illumina platform) was developed for construction of NGS libraries for methylated CpG sites using bisulfite treated DNA (20 ng – 500 ng) as input. The kit enriches methylated CpG regions, thus significantly reduce the sequencing cost. The kit estimates the whole genome methylation patterns at the single base level since it is based on a bisulfite-seq technology.
It is known that bisulfite treatment of completed NGS libraries causes tremendous damage to the libraries. By using bisulfite treated DNA as input, the kit overcomes the significant library loss due to the bisulfite conversion. The kit contains a mixture of PCR polymerases that have high-fidelity amplification and uracil tolerance which is ideal for bisulfite treated DNA.
BioDynami Methylation Specific Bisulfite Seq建库试剂盒工作流程
试剂盒有三种index类型:
Non-index(Cat.# 30101):没有索引。
Index(Cat.# 30102):每个引物包含6个碱基的唯一序列,用于识别单个库。库复用能力可达48个样本。
Unique dual index(Cat.# 30103):重亚硫酸氢盐测序文库复用多达96个样本,具有独特的双指标。BioDynami使用了一个Four-Base Difference Index System来生成在8基指数中至少有4个不同的指数。索引引物消除了NGS的错误,包括索引交叉污染、索引跳跃、读取错误赋值等。
试剂盒优势:
★ 甲基化CpG位点的富集
★ 单碱基分辨率
★ 测序成本低
★ 迅速
★ 工作流程简单
★ 亚硫酸氢盐处理的DNA作为输入:从20 ng到500 ng
★ 总时间:1.5小时
★ 操作时间:10分钟
MSBS文库准备试剂盒丰富了CpG位点
人类基因组中的高甲基化区域和低甲基化区域
人类基因组中的高甲基化区域
人类基因组中的低甲基化区域
测序设置:推荐单端35个周期(读取1,35个碱基)
为了最大限度地提高甲基化CpG的富集,我们建议用单末端35个循环(read1,35个碱基)对MSBS文库进行测序。这是因为富集的甲基化CpG位点主要位于read 1序列的开头附近。单端reads越短,甲基化CpG富集越好。
产品订购:
产品名称 |
货号 |
产品规格 |
Methylation Specific Bisulfite Seq Library Prep Kit |
30101S |
non-index,24 reactions |
Methylation Specific Bisulfite Seq Library Prep Kit |
30101L |
non-index,48 reactions |
Methylation Specific Bisulfite Seq Library Prep Kit |
30102S |
index,24 reactions |
Methylation Specific Bisulfite Seq Library Prep Kit |
30102L |
index,48 reactions |
Methylation Specific Bisulfite Seq Library Prep Kit |
30103S |
unique dual index,96 reactions |
Methylation Specific Bisulfite Seq Library Prep Kit |
30103L |
unique dual index,192 reactions |
风险提示:丁香通仅作为第三方平台,为商家信息发布提供平台空间。用户咨询产品时请注意保护个人信息及财产安全,合理判断,谨慎选购商品,商家和用户对交易行为负责。对于医疗器械类产品,请先查证核实企业经营资质和医疗器械产品注册证情况。