EpiTect Plus Bisulfite Kits

对多种起始材料进行DNA亚硫酸氢盐转化，无需单独的DNA纯化步骤

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EpiTect Plus DNA Bisulfite Kit

Cat. No. / ID: 59124

48 MinElute DNA spin columns, Bisulfite mix, DNA Protect Buffer, Carrier RNA, Buffers

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EpiTect Plus Bisulfite Kits 旨在用于分子生物学应用。这些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

适用于基因组DNA、全血、细胞、组织和FFPE组织切片
高效进行样本裂解和亚硫酸氢盐转化
产物为即用型DNA，产量高，洗脱体积小
独特的DNA保护技术可防止DNA降解
最大程度上减少手工操作，快速获得结果

Product Details

EpiTect Plus Bisulfite Kits是一组整合的试剂盒体系，适用于各种起始材料，可高效的进行样本裂解，直接对从全血、培养细胞、组织样本和FFPE组织切片中提取的DNA进行亚硫酸氢盐转化。转化后的DNA可洗脱到10 µl的洗脱缓冲液中，因此DNA的浓度高。EpiTect Plus Bisulfite Kits还具有同样高的DNA转化效率，以及EpiTect Bisulfite Kits可靠的DNA保护技术。

Performance

使用EpiTect Plus Bisulfite Kits得到的DNA的产量是其他试剂盒的4–8倍（参见""和""）。产量增加，使得下游操作更加灵活；因为由一个样本得到的转化后DNA可用于多个后续操作，或储存以备将来使用。目前QIAGEN的数据显示：使用EpiTect Plus Bisulfite Kits转化和纯化的DNA可稳定储存至少3年（参见""）。

See figures

High recovery of converted DNA.

Superior yields.

DNA storage stability.

Principle

EpiTect Plus Bisulfite Kits的主要特点包括：

DNA的转化效率超过99%，且DNA的降解程度达到最小
转化后的DNA浓度高，洗脱体积小（低至10 µl）
整合的技术可最大程度上减少样本裂解过程中的DNA损失和保护DNA，转化效率高
优化、简便的操作流程适用于基因组DNA、全血、培养细胞、组织样本和FFPE组织切片
起始材料用量灵活：1 ng–2 µg gDNA、0.5–20 µl全血、10–100,000个细胞或1片FFPE组织切片（10 µm厚）
可进行组织裂解，节省时间

使用亚硫酸氢钠进行DNA转化时，所有样本DNA中非甲基化的胞嘧啶都被完全转化，同时减少了由于温度和化学试剂因素导致的DNA降解。对于转化后DNA的保护十分重要，影响回收率和下游操作的灵敏度。EpiTect Plus Bisulfite Kits在防止DNA降解的同时，可高效的进行DNA亚硫酸氢盐转化，且无需单独的DNA纯化步骤。这个高效的工作流程覆盖了从样本裂解到最终洗脱转化后DNA的整个过程，可快速获得适合多种下游应用的高品质、即用型DNA。

QIAGEN对DNA亚硫酸氢盐转化的解决方案—找到适合您的试剂盒

3种EpiTect Plus Bisulfite Kits相互关联，是一个整体，这3种试剂盒可对多种起始材料的DNA进行直接的亚硫酸氢盐转化（参见"Interconnected protocols"）。但每种试剂盒有自己独特的特点，适用于某一特定样本类型，含有进行特定转化所需的所有缓冲液和试剂。使用每个试剂盒所含的试剂可进行高效的样本裂解，裂解过程的DNA损失降到最小；完全的亚硫酸氢盐转化，转化过程中有效的保护DNA和彻底回收DNA。

EpiTect Plus Bisulfite Kits不能替代EpiTect Bisulfite Kits，但适用于更多样本类型，并可为下游应用制备亚硫酸氢盐转化DNA。两套试剂盒都有DNA保护技术，可确保高转化效率。EpiTect Bisulfite Kits提供便利的96孔板，适合高通量转化。下表"QIAGEN对DNA亚硫酸氢盐转化的解决方案"中列出了QIAGEN表观遗传学产品线中的5个亚硫酸氢盐转化试剂盒，以便您从中选择合适的试剂盒。

整合的技术—DNA Protect Buffer和Deparaffinization Solution

独特配方的DNA Protect Buffer，可预防在高温、低pH环境下对DNA进行亚硫酸氢盐处理时产生的DNA片段化。该缓冲液可促使DNA变性，生成胞嘧啶完全转化所需的单链DNA。DNA Protect Buffer含有pH指示染料，可确保胞嘧啶转化时pH准确（参见" DNA Protect Buffer"）。

脱蜡过程可将DNA从FFPE样本中提取出来。在该过程中，石蜡先被溶解，然后去除；再用裂解缓冲液和蛋白酶K处理样本。Deparaffinization Solution是新型试剂，可减少去石蜡化过程所需的手动操作，并可确保高DNA回收率。在溶液中加入FFPE样本后，溶液作用于样本，同时蛋白酶K开始消化。之后无需离心沉淀FFPE样本，也无需去除含有石蜡的上清。因此避免了去石蜡化过程中的样本损失。

QIAGEN对DNA亚硫酸氢盐转化的解决方案
特点	EpiTect Bisulfite Kit	EpiTect 96 Bisulfite Kit	EpiTect Plus DNA Bisulfite Kit	EpiTect Plus LyseAll Bisulfite Kit	EpiTect Plus FFPE Bisulfite Kit
规格	EpiTect离心柱	96孔板	MinElute DNA离心柱	MinElute DNA离心柱	MinElute DNA离心柱
起始材料	gDNA	gDNA	gDNA	全血、细胞、组织	FFPE组织切片
起始材料用量	1 ng – 2 µg gDNA	1 ng – 2 µg gDNA	1 ng – 2 µg gDNA	10–100,000个细胞，0.5–20 µl全血	1片FFPE组织切片（10 µm厚）
是否包括样本裂解	否	否	否	是，含有可裂解细胞、血液和组织的Lysis Buffer FTB	是，含有Deparaffinization Solution和Lysis Buffer FTB
洗脱体积	20–40 µl	40–50 µl	≥10 µl	≥10 µl	≥10 µl
转化效率	>99%	>99%	>99%	>99%	>99%
自动化	QIAcube全自动核酸纯化仪	无	QIAcube全自动核酸纯化仪	QIAcube全自动核酸纯化仪	QIAcube全自动核酸纯化仪
离心/真空	离心	离心/真空	离心	离心	离心
试剂盒组成	EpiTect Bisulfite Kit	EpiTect 96 Bisulfite Kit	EpiTect Plus DNA Bisulfite Kit	EpiTect Plus LyseAll Lysis Kit和EpiTect Plus DNA Bisulfite Kit	EpiTect Plus FFPE Lysis Kit和EpiTect Plus DNA Bisulfite Kit

See figures

DNA Protect Buffer.

Procedure

EpiTect Plus DNA Bisulfite Kit

EpiTect Plus DNA Bisulfite Kit含有特别配制的缓冲液和试剂，可对基因组DNA进行高效的亚硫酸氢盐转化和回收（参见" EpiTect Plus DNA flowchart"）。该试剂盒具有独特的DNA保护技术和QIAGEN离心柱，可确保完全转化DNA、降解程度最低，且可洗脱到10 µl的缓冲液中，适合需要高浓度DNA的下游操作。该试剂盒可用于从20 µl溶液中转化1 ng–2 µg DNA或40 µl溶液中转化1–500 ng DNA。

EpiTect Plus FFPE Bisulfite Kit

EpiTect Plus FFPE Bisulfite Kit包括用于去除石蜡和裂解FFPE组织切片的EpiTect Plus FFPE Lysis Kit和用于对样本DNA进行高效亚硫酸氢盐转化的EpiTect Plus DNA Bisulfite Kit（参见" EpiTect Plus FFPE flowchart"）。使用新型的Deparaffinization Solution和Lysis Buffer FTB，在优化的温度下，对FFPE组织切片进行去石蜡化、裂解和去交联操作。裂解和去交联后的DNA可直接用EpiTect Plus DNA Bisulfite Kit进行亚硫酸氢盐转化。该试剂盒的起始材料为厚度为10 µm的单片FFPE组织切片。

EpiTect Plus LyseAll Bisulfite Kit

EpiTect Plus LyseAll Bisulfite Kit包括用于裂解全血细胞、培养细胞和新鲜组织的EpiTect Plus LyseAll Lysis Kit和用于对样本DNA进行高效亚硫酸氢盐转化的EpiTect Plus DNA Bisulftie Kit（参见" EpiTect Plus LyseAll flowchart"）。特别配制的裂解缓冲液可更高效的进行组织裂解，得到的DNA可直接用于亚硫酸氢盐转化，无需单独的纯化步骤。该试剂盒适用于0.5–20 µl血液或10–10⁵个细胞（至少60 pg DNA）。

See figures

EpiTect Plus DNA flowchart.

EpiTect Plus FFPE flowchart.

EpiTect Plus LyseAll flowchart.

Applications

使用EpiTect Plus Bisulfite Kit转化和纯化的DNA适用于多种下游应用，包括：

甲基化特异性PCR
多重PCR
Real-time PCR
测序/焦磷酸测序

Supporting data and figures

Interconnected protocols of the EpiTect Fast 96 Bisulfite Conversion Kits.

Depending on sample type, the bisulfite conversion of DNA calls for the use of a specific kit and protocol. Kits contain buffers optimized for DNA extraction and all reagents for the DNA bisulfite conversion.

Resources

Download Safety Data Sheets for QIAGEN product components.

FAQ

The MinElute spin columns included in the following kits should be stored at 2–8°C upon arrival: AllPrep DNA/RNA Micro, EpiTect Fast DNA Bisulfite, EpiTect Fast FFPE Bisulfite, EpiTect Fast LyseAll Bisulfite, EpiTect Plus DNA Bisulfite, EpiTect Plus FFPE Bisulfite, EpiTect Plus LyseAll Bisulfite, exoRNeasy Serum/plasma Maxi, exoRNeasy Serum/Plasma Midi, GeneRead DNA FFPE, GeneRead rRNA Depletion, GeneRead Size Selection, MinElute Gel Extraction, MinElute PCR Purification, MinElute Reaction Cleanup, miRNeasy FFPE, miRNeasy Micro, miRNeasy Serum/Plasma, QIAamp DNA FFPE, QIAamp DNA Investigator, QIAamp DNA Micro, QIAamp MinElute Media, QIAamp MinElute Virus Spin, QIAamp MinElute Virus Vacuum, RNeasy FFPE, RNeasy Micro, RNeasy Plus Micro.

Short-term storage (up to 4 weeks) at room temperature (15–25°C) does not affect the performance. However, for optimal performance and quality, storage temperature should not exceed 25°C.

FAQ ID - 3560

The bisulfite conversion chemistry of the EpiTect Plus Kits remain unchanged compared to the classic EpiTect Bisulfite Conversion Kit. The only change made in the new kit has been the new columns which allow for a smaller elution volume. Since the Bisulfite conversion chemistry part is responsible for the Bisulfite conversion, use of the new kit does not have an effect on the conversion rate. The EpiTect Plus Kits benefit from a smaller elution volume allowing it to be paired with methods that have less sample and allow direct isolation for cultured cells and blood cells using the LyseAll Bisulfite kit. Another application is direct isolation and conversion of FFPE samples using the EpiTect Plus FFPE Lysis kit. These isolate DNA directly from samples and bisulfite convert them before prior to isolation.

FAQ - 3344

Yes, DNA converted with EpiTect Bisulfite Kits and EpiTect Plus Bisulfite Kits is very stable and can be stored at -20 C for at least 3 years without decrease in quality or conversion. See the website for more details and data: http://www.qiagen.com/products/epigenetics/epitect/epitectbisulfitekit.aspx#Tabs=t1

FAQ ID -2409

The typical yields of bisulfite converted DNA when using the EpiTect Plus Bisulfite Kits depend on the amount of DNA and source of the starting material. Typically over 80% of DNA can be recovered after bisulfite conversion using EpiTect Plus DNA Kit.

Input amounts are as follows:

DNA: 1ng-2µg (EpiTect Plus DNA Bisulfite Kit)

Cells: 10-10E5 (EpiTect Plus LyseAll Bisulfite Kit)

Blood:0.5-20 µl (EpiTect Plus LyseAll Bisulfite Kit)

FFPE tissue:1 slice (10µm thick) (EpiTect Plus FFPE Bisulfite Kit)

FAQ ID -2408

The DNA Protect buffer included in all EpiTect Bisulfite and EpiTect Plus Bisulfite kits works in two ways.

1) it serves as a scavenger for free radicals which are built during bisulfite conversion. As these free radicals can harm and fragment DNA the scavenger protects DNA from excessive degradation.

2) it keeps DNA single stranded even at lower temperatures so that DNA doesn’t need to be subjected to high temperatures for long time which is known to degrade DNA. Thus, it ensures high conversion rates even at gentle conversion conditions.

This is of special interest if working with low DNA qualities e.g. when working with formalin-fixed paraffin-embedded tissue samples.

FAQ ID -2410

The EpiTect Plus LyseAll Bisulfite Kit is recommended for whole blood as well as for cells. The kit includes an innovative lysis buffer for optimized lysis of up to 10E5 cells and up to 20µl whole blood. It also facilitates binding of the DNA to ensure that even small amounts of DNA are bound with high efficiency, thus reducing loss of DNA due to the DNA purification step.

For tissues:e

A) For fresh or frozen tissue the EpiTect Plus LyseAll Bisulfite Kit should be used. The kit includes an innovative lysis buffer for optimized lysis of tissue samples. It also facilitates binding of the DNA to ensure that even small amounts of DNA are bound with high efficiency, thus reducing loss of DNA due to the DNA purification step.

B) For FFPE tissue we strongly recommend the EpiTect Plus FFPE Bisulfite Kit. It includes specialized buffers for efficient deparaffinization and lysis of FFPE tissue samples. After lysis DNA is directly bisulfite conversion without prior purification of DNA thus eliminating loss of DNA due to DNA isolation.

FAQ ID -2411

No. Under the current protocol, FFPE samples are not suitable for analysis with EpiTect Methyl qPCR Arrays and Assays. This is because genomic DNA from FFPE samples is single-stranded due to the heating step involved in the DNA isolation process, making it undigestible by either Methylation-sensitive or Methylation-dependent enzymes. Please refer to the User Manual of your chosen genomic DNA isolation kit. If there is a 95 ºC heating step involved, please do not use the kit to isolate genomic DNA for EpiTect Methyl qPCR Arrays or Assays.

Instead, use the recommended genomic DNA isolation Kit in the User Manual of EpiTect Methyl qPCR Arrays or Assays. For analyzing methylation status of DNA isolated from FFPE tissues, we recommend using EpiTect Plus FFPE Bisulfite Kits for one-step isolation and bisulfite conversion of the DNA, followed by any detection method optimized for use with FFPE samples (e.g., Pyrosequencing).

FAQ ID -2733

The difference in these two kits are the spin columns. EpiTect Plus Bisulfite Kit contains the MinElute DNA Spin Columns and the EpiTect Bisulfite Kit contains the EpiTect Spin columns. The MinElute Spin Columns in the EpiTect Plus Bisulfite Kit have a lower elution volume, lower dead volume and lower absolute binding capacity. The binding capacities for both columns are far higher than what you put into your conversion reaction. The conversion rate and recovery rate for both kits are the same.

FAQ ID -3167

The EpiTect Plus FFPE Bisulfite Kit includes specialized buffers for efficient deparaffinization and lysis of FFPE tissue samples. After lysis DNA is directly bisulfite conversion without prior purification of DNA thus eliminating loss of DNA due to DNA isolation. In addition, the DNA Protect buffer included in the kit minimizes the DNA fragmentation due to bisulfite treatment.

In this combination, the yield of analyzable DNA can be increased 200 fold. A 25 fold due to direct isolation and 8-9 fold by DNA protection.

FAQ ID -2412