EpiTect Plus Bisulfite Kits

For complete DNA bisulfite conversion from various starting materials, without prior DNA isolation

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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

EpiTect Plus Bisulfite Kits están concebidos para su uso en aplicaciones de biología molecular. Estos productos no están concebidos para el diagnóstico, la prevención ni el tratamiento de enfermedades.

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

Features

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

Product Details

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.

Performance

DNA output from EpiTect Plus Bisulfite Kits is 4–8 times greater compared with kits tested from other suppliers (see figure "" and ""). 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 "").

See figures

High recovery of converted DNA.

Superior yields.

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

See figures

DNA Protect Buffer.

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).

See figures

EpiTect Plus DNA flowchart.

EpiTect Plus FFPE flowchart.

EpiTect Plus LyseAll flowchart.

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

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.

Cleanup of 100 ng or more bisulfite converted DNA

This protocol is for cleanup of 100 ng or more bisulfite converted DNA using the EpiTect Plus DNA Bisulfite Kit.

Sample Size	140 µl bisulfite reaction
Elution volume	10-16 µl in increments of 1 µl; default 16 µl
Applications	Cleanup, DNA
Starting material	Bisulfite converted DNA, Bisulfite converted DNA derived from FFPE tissues

Cleanup of 100 ng or more bisulfite converted DNA

This protocol is for cleanup of up to 100 ng bisulfite converted DNA using the EpiTect Plus DNA Bisulfite Kit.

Sample Size	140 µl bisulfite reaction
Elution volume	10-16 µl in increments of 1 µl; default 16 µl
Applications	Cleanup, DNA
Starting material	Bisulfite converted DNA, Bisulfite converted DNA derived from FFPE tissues

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