EpiTect Plus Bisulfite Kits

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 DNAwhile 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

The3EpiTect 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 ofthe 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.

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).

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