✓ 24/7 automatic processing of online orders
✓ Knowledgeable and professional Product & Technical Support
✓ Fast and reliable (re)-ordering
EpiTect Whole Bisulfitome Kit (25)
Cat. No. / ID: 59203
REPLI-g Midi DNA Polymerase, EpiTect WBA Reaction Buffer, nuclease-free water for 25 whole bisulfitome amplification reactions
Log in To see your account pricing.
EpiTect Whole Bisulfitome Kit 旨在用于分子生物学应用。该产品不能用于疾病诊断、预防和治疗。
✓ 24/7 automatic processing of online orders
✓ Knowledgeable and professional Product & Technical Support
✓ Fast and reliable (re)-ordering
Features
- 获得高产量的亚硫酸氢盐转化DNA，可用于多个PCR
- 专用于扩增亚硫酸氢盐转化后DNA
- 方便的一步建立反应体系
Product Details
EpiTect Whole Bisulfitome Kit包含DNA聚合酶、缓冲液和试剂，采用多重置换扩增（MDA）技术，用于经亚硫酸氢盐转化后DNA的全基因组扩增。EpiTect Whole Bisulfitome Kit使用经验证的REPLI-g技术，专为扩增经亚硫酸氢盐转化DNA研发（亚硫酸氢盐转化后其核苷酸组成已发生改变，且DNA片段更小），同时维持转化后序列。EpiTect Whole Bisulfitome Kit的常规DNA产量是每次反应1–3 μg，因此，单次亚硫酸氢盐反应获得的模板DNA可进行多达300次PCR分析。
Performance
EpiTect Whole Bisulfitome Kit是为所有甲基化分析提供足够DNA的理想工具，而不受亚硫酸氢盐转化后DNA量较少的限制。亚硫酸氢盐转化后的全基因组扩增（WBA）流程可获得高产的亚硫酸氢盐转化后DNA，同时配合含有独特DNA保护体系的EpiTect Bisulfite Kits维持转化后序列的表现。EpiTect Whole Bisulfitome Kit经检测优于其他供应商提供的扩增试剂盒（参见" High amplification efficiency"和" Reliable whole bisulfitome amplification"）。
See figures
Principle
基因组DNA序列的甲基化分析正面临着挑战，尤其在样本量较少的情况下。使用EpiTect Whole Bisulfitome Kit可高度一致的扩增亚硫酸氢盐转化后DNA的全基因组，无序列偏向性（参见" EpiTYPER MALDI-TOF analysis of WBA DNA"）。该方法基于等温多重置换扩增（MDA）技术。复制过程中通过带有3'→5'外切酶校正活性的行进性DNA聚合酶确保其高保真扩增。
EpiTect Whole Bisulfitome Kit应用经验证的REPLI-g技术，专为扩增经亚硫酸氢盐转化后DNA研发（亚硫酸氢盐转化后其核苷酸组成已发生改变，且DNA片段更小），同时维持转化后序列的表现。配合含有独特DNA保护体系的EpiTect Bisulfite Kits，可获得最佳结果。这样可确保DNA有效变性，生成胞嘧啶完全转化所必需的单链DNA，同时避免由于高温、低pH值的亚硫酸氢盐处理引起的DNA过度断裂。通过预防DNA断裂可确保后续较长DNA片段的扩增。使用EpiTect Whole Bisulfitome Kit，通常每个反应获得的DNA的量为1–3 μg。
See figures
Procedure
方便的一步反应体系，即使用反应缓冲液和REPLI-g DNA聚合酶的混合液孵育亚硫酸氢盐转化后的模板DNA。加热失活聚合酶后，扩增的DNA可直接进行分析或贮存（参见" WBA procedure"）。
生物学和医药研究的很多领域中，尤其是肿瘤学、干细胞研究和发育生物学研究中，获得表观遗传学信息是首要任务。但是，由于缺乏提供可重复性数据的标准化方法，尤其是样本材料的限制，分析DNA的甲基化变化非常困难。QIAGEN最新推出的EpiTect方案，建立了从DNA样本收集、稳定和纯化到亚硫酸氢盐转化和real-time或终点式PCR甲基化分析或测序的标准化预分析和分析方案（参见" Standardized workflows in epigenetics"）。
See figures
Applications
EpiTect Whole Bisulfitome Kit是为所有甲基化分析提供足够DNA的理想工具，而不受亚硫酸氢盐转化后DNA量较少的限制。每次使用10 ng DNA模板扩增获得的DNA，可进行100–300次real-time PCR分析（如使用EpiTect MethyLight PCR Kits）或终点法PCR（如使用EpiTect MSP Kits）。
Supporting data and figures
Reliable whole bisulfitome amplification.
Real-time PCR was used to measure the representation of four different loci in 10 ng bisulfite converted DNA, before (–WBA) and after (+WBA) amplification with the EpiTect Whole Bisulfitome Kit. DNA was first bisulfite converted using either the EpiTect Bisulfite Kit or a kit from supplier Z, and was then amplified; qPCR was performed using the QuantiTect SYBR® Green PCR Kit. In the EpiTect converted DNA, the four loci were represented similarly before and after amplification. Higher CT values for the DNA converted with the competitor’s bisulfite kit indicate higher DNA fragmentation prior to WBA. Following amplification, the relative representation of the four loci was increased and different, indicating lower whole bisulfitome amplification and unequal amplification.
Specifications
| Features | Specifications |
|---|---|
| Amplification | Whole bisulfite converted DNA |
| Reaction time | 8h at 28°C |
| Maximum input volume | 10 µl bisulfite converted DNA |
| Starting amount of DNA | > 50 ng bisulfite converted DNA |
| Applications | Methylation specific PCR |
| Reaction volume | 40 µl |
| Starting material | Epitect bisulfite converted DNA |
| Technology | Multiple Displacement Amplification (MDA) |
| Yield | 1-3 µg per reaction |
Resources
试剂盒操作手册 (1)
安全数据表 (1)
Safety Data Sheets (1)
Certificates of Analysis (1)
Kit Handbooks (1)
FAQ
How long can DNA converted with EpiTect Bisulfite Kits be stored before use in methylation specific PCR with the EpiTect MSP Kit?
3318 - What is the preferred amplicon size for performing PCR on DNA amplified with the EpiTect Whole Bisulfitome Kit?