For complete DNA bisulfite conversion from various starting materials, without prior DNA isolation
  • Dedicated solutions for gDNA, blood, cells, tissues, and FFPE samples
  • Streamlined procedure combining sample lysis and bisulfite conversion
  • High-yield recovery of ready-to-use DNA in small elution volumes
  • Unique DNA Protect Technology to prevent DNA degradation
  • Minimal handling and rapid results
The EpiTect Plus Bisulfite Kits are a coordinated system of kits that offer exceptional flexibility in starting material and a streamlined procedure for sample lysis and direct bisulfite conversion of DNA released from whole blood, cell cultures, tissue samples, and FFPE tissue slices. In addition, the converted DNA can be eluted in as little as 10 µl elution buffer, resulting in high DNA concentration. EpiTect Plus Bisulfite Kits continue to feature the same high DNA conversion rate and reliable DNA Protect Technology of the EpiTect Bisulfite Kits. Procedures can be automated on the QIAcube.
 
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EpiTect Plus DNA Bisulfite Kit
48 MinElute DNA spin columns, Bisulfite mix, DNA Protect Buffer, Carrier RNA, Buffers
59124
214,00 €

EpiTect Plus Bisulfite Kits are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.


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DNA storage stability.|DNA Protect Buffer.|Superior yields.|High recovery of converted DNA.|EpiTect Plus DNA flowchart.|EpiTect Plus LyseAll flowchart.|EpiTect Plus FFPE flowchart.|Interconnected protocols.|
Bisulfite converted DNA generated with EpiTect Kits was stored at –20°C for up to 36 months, and amplified at several time intervals. The resulting threshold cycle values at all time points for various DNA amounts were comparable to those obtained with freshly converted DNA (time point 0).|[A] Upon addition to the bisulfite conversion reaction, the DNA Protect Buffer [B] should turn from green to blue, indicating sufficient mixing and correct pH.|DNA was purified from rat liver FFPE tissue (10 µm) using the EpiTect Plus FFPE Bisulfite Kit (1–4) or an alternative kit from Supplier Z (5–8). [A] Detection of the indicated genes using the PyroMark PCR Kit and agarose gel analysis. [B] Detection of two amplicons of the GSTP gene (156 and 279 bp) using the QuantiTect SYBR® Green PCR Kit (4 triplicate samples). Templates purified using the EpiTect Plus FFPE Bisulfite Kit resulted in lower CT values and highly reproducible results for both amplicons (left). Templates purified using the kit from Supplier Z resulted in amplification of only the 156 bp fragment with higher CT values (right).|Degradation of DNA during the bisulfite conversion reaction results in low yields of the target sequence for downstream applications. This problem is particularly important for samples such as FFPE tissues, where the starting DNA is already fragmented. The DNA Protect Buffer included in EpiTect Plus Bisulfite Kits minimizes DNA damage during conversion. Along with minimal loss of DNA during sample lysis and thorough cleanup of the resulting converted DNA, this innovative DNA protect technology maximizes recovery of the converted DNA.|Streamlined and straightforward, the EpiTect Plus DNA Bisulfite Kit enables complete bisulfite conversion and clean up of genomic DNA in approximately 6 hours.|Upon completion of the sample lysis steps, bisulfite conversion is completed using the included EpiTect Plus DNA Bisulfite Kit.|Upon completing deparaffinization, lysis, and decrosslinking steps, bisulfite conversion is completed using the EpiTect Plus DNA Bisulfite Kit.|Depending on sample type, the bisulfite conversion of DNA calls for the use of a specific kit and protocol. EpiTect Plus Bisulfite Kits contain buffers optimized for DNA extraction and all reagnets for the DNA bisulfite conversion.|
Performance
DNA output from EpiTect Plus Bisulfite Kits is 4–8 times greater compared with kits tested from other suppliers (see figure "High recovery of converted DNA" and "Superior yields"). Greater DNA output translates to greater flexibility in downstream applications, as DNA from one sample can be subjected to a greater number of subsequent investigations or stored for future use. Current data at QIAGEN indicate that DNA converted and purified with EpiTect Plus Bisulfite Kits is stable for at least 3 years (see figure "DNA storage stability").
Principle

Key features of the EpiTect Plus Bisulfite Kits include:

Over 99% DNA conversion rate with minimal degradation
High concentration of converted DNA in low elution volumes (down to 10 µl)
Integrated technologies to minimize DNA loss during sample lysis and protect DNA, for unparalleled yields
Convenient, carefully optimized protocols for gDNA, whole blood, cell cultures, tissue samples, and FFPE
High input range from 1 ng – 2 µg gDNA, 0.5–20 µl whole blood, 10–100,000 cells, or 1 FFPE tissue slice (10 µm thick)
Integrated lysis procedure and streamlined protocols to save time

Effective conversion of DNA by treatment with sodium bisulfite must accomplish complete conversion of all unmethylated cytosines in the sample DNA while minimizing DNA degradation due to the harsh temperature and chemical conditions of the bisulfite conversion reaction. The protection of the treated DNA is a critical factor in the recovery rate of converted DNA and in the sensitivity of downstream applications. EpiTect Plus Bisulfite Kits facilitate effective bisulfite conversion of DNA while minimizing DNA degradation, with the added advantage of not requiring DNA isolation prior to the conversion reaction. This streamlines the workflow from sample lysis to final elution of converted DNA, providing rapid results and high-quality DNA ready to use in a variety of downstream applications.

QIAGEN solutions for DNA bisulfite conversion — the right kit for your needs

The 3 EpiTect Plus Bisulfite Kits are interconnected, functioning as an integrated system of solutions for direct bisulfite conversion of DNA from a broad range of starting materials (see figure "Interconnected protocols"). However, each kit has unique features, addresses a specific sample type, and includes all buffers and reagents necessary to function independently. The components in each kit combine to deliver effective lysis of samples, minimal loss of DNA during lysis, complete bisulfite conversion, robust protection during conversion, and thorough cleanup of DNA. 

The EpiTect Plus Bisulfite Kits do not replace the EpiTect Bisulfite Kits, but rather expand the types of samples that can be prepared for downstream applications requiring bisulfite converted DNA, and integrate the DNA preparation procedure. Both sets of kits include DNA Protect Technology and accomplish high conversion rates. EpiTect Bisulfite Kits offer a convenient 96-well format for high sample throughput. See table "QIAGEN solutions for DNA bisulfite conversion" for an overview of all 5 bisulfite conversion solutions in QIAGEN's Epigenetics portfolio to easily select the best kit for your application.

Integrated technologies — DNA Protect Buffer and Deparaffinization Solution

DNA Protect Buffer is uniquely formulated to prevent the fragmentation usually associated with bisulfite treatment of DNA at high temperatures and low pH values. It also provides effective DNA denaturation, resulting in the single-stranded DNA necessary for complete cytosine conversion. DNA Protect Buffer contains a pH indicator dye as a mixing control that allows confirmation of the correct pH for cytosine conversion (see figure "DNA Protect Buffer").

To extract DNA from FFPE samples, deparaffinization needs to be carried out. This is a process whereby paraffin is first dissolved and then removed, exposing samples for subsequent treatment with lysis buffer and proteinase K. Deparaffinization Solution is an innovative chemistry that facilitates deparaffinization with minimal handling. A great advantage of using Deparaffinization Solution is that it contributes to high DNA recovery rates. After addition to an FFPE sample, the solution remains on the sample while proteinase K digestion is carried out. There is no need to pellet the FFPE sample thereafter, or to remove paraffin-containing supernatant. Thus, the risk of sample loss during deparaffinization is avoided.

QIAGEN solutions for DNA bisulfite conversion
Feature EpiTect Bisulfite Kit EpiTect 96 Bisulfite Kit EpiTect Plus DNA Bisulfite Kit EpiTect Plus LyseAll Bisulfite Kit  EpiTect Plus FFPE Bisulfite Kit
Format EpiTect spin column 96-well plate MinElute DNA spin column MinElute DNA spin column MinElute DNA spin column
Starting material gDNA gDNA gDNA Whole blood, cells, tissues FFPE tissue slices
Input amount 1 ng – 2 µg gDNA 1 ng – 2 µg gDNA 1 ng – 2 µg gDNA 10–100,000 cells, 0.5–20 µl whole blood 1 FFPE tissue slice (10 µm)
Sample lysis step included?  No  No  No Yes. Lysis Buffer FTB for cells, blood, and tissues is included Yes. Deparaffinization Solution and Lysis Buffer FTB are included
Elution volume 20–40 µl 40–50 µl ≥10 µl ≥10 µl ≥10 µl
Conversion efficiency >99% >99% >99% >99% >99%
Automation QIAcube No QIAcube QIAcube QIAcube
Centrifugation/vacuum protocol Centrifugation Centrifugation/vacuum Centrifugation Centrifugation Centrifugation
Kit composition EpiTect Bisulfite Kit EpiTect 96 Bisulfite Kit EpiTect Plus DNA Bisulfite Kit EpiTect Plus LyseAll Lysis Kit and EpiTect Plus DNA Bisulfite Kit EpiTect Plus FFPE Lysis Kit and EpiTect Plus DNA Bisulfite Kit

Procedure
EpiTect Plus DNA Bisulfite Kit

The EpiTect Plus DNA Bisulfite Kit includes carefully formulated buffers and reagents to provide streamlined and effective bisulfite conversion of genomic DNA and subsequent cleanup of the converted DNA (see figure "EpiTect Plus DNA flowchart"). With unique DNA Protect Technology and QIAGEN spin columns, this kit generates fully converted DNA with minimal degradation that can be eluted in as little as 10 µl of buffer for downstream applications requiring high DNA concentrations. This kit can be used for conversion of 1 ng – 2 µg DNA in a volume of 20 µl or 1–500 ng DNA in a volume of 40 µl.

EpiTect Plus FFPE Bisulfite Kit

The EpiTect Plus FFPE Bisulfite Kit consists of the EpiTect Plus FFPE Lysis Kit for the deparaffinization and lysis of FFPE tissue slices, and the EpiTect Plus DNA Bisulfite Kit for the effective bisulfite conversion of the DNA released from the samples (see figure "EpiTect Plus FFPE flowchart". The deparaffinization, lysis, and decrosslinking of FFPE slices is optimized with innovative Deparaffinization Solution and Lysis Buffer FTB, and a carefully designed reaction temperature protocol. The DNA released during lysis and decrosslinking is used directly for bisulfite conversion with the included EpiTect Plus DNA Bisulfite Kit. Input amount for this kit is single FFPE tissue slices of 10 µm in thickness.

EpiTect Plus LyseAll Bisulfite Kit

The EpiTect Plus LyseAll Bisulfite Kit consists of the EpiTect Plus LyseAll Lysis Kit for lysis of whole blood cells, cells from culture, and fresh tissues, and the EpiTect Plus DNA Bisulftie Kit for the effective bisulfite conversion of the DNA released from these samples (see figure "EpiTect Plus LyseAll flowchart". Uniquely formulated lysis buffers streamline the release of DNA from samples, which can be used directly for bisulfite conversion without the need for prior isolation. This kit can be used with 0.5–20 µl blood or 10–105 cells (as little as 60 pg of DNA).

Applications

DNA converted and purified using EpiTect Plus Bisulfite Kits is suitable for use in a range of downstream applications including:

Methylation-specific PCR
Multiplex PCR
Real-time PCR
Sequencing/Pyrosequencing

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Kit Handbooks
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メチル化解析のため最適化されたBisulfite 法による完全なDNA 変換およびクリーンアップFFPE 組織、血液、培養細胞、組織サンプルから
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