Cells-to-CpG™ Bisulfite Conversion & Quantitation Control Kit

Catalog number: 4445553

Applied Biosystems™  Related applications: Epigenetic Sequencing | Methylation Analysis | SOLiD® Next-Generation Sequencing

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Description

When performing methylation studies using the Cells-to-CpG™ Bisulfite Conversion Kit, use the components of the Cells-to-CpG™ Bisulfite Conversion and Quantitation Control Kit to serve as controls during bisulfite conversion and subsequent amplification of the bisulfite-converted DNA. The included Unconverted Human Male gDNA (Control Sample 1) (100 ng/ µL) is used as an external sample control during the bisulfite conversion, and the Converted Human Male gDNA (Control Sample 2) (10 ng/ µL) is used to quantify the amount of bisulfite-converted human genomic DNA in your samples.

The bisulfite method is the most commonly used technique for identifying specific methylation patterns within a DNA sample. It consists of treating DNA with bisulfite, which converts unmethylated cytosines to uracil but does not change methylated cytosines (Figure 1). Bisulfite conversion has been utilized in DNA methylation research for the last 20 years with few improvements to the technology until now. With thorough optimization, the Cells-to-CpG™ Bisulfite Conversion Kit provides a quick, streamlined method (Figure 2) for bisulfite conversion to reveal methylated cytosines in either loci-specific or genomewide analyses.

Key product features:
• Flexible DNA conversion—validated solution for our platforms to minimize the need for optimization
• Quantitative reliability—confidence in results with available controls to easily monitor conversion rate
• Streamlined workflow—ability to start with various sample input types including cell, tissue, blood, and FFPE samples
• Efficient application—reduced time and labor through direct conversion of cytosines in samples without need for purifying genomic DNA

Complete DNA Conversion with Greater Flexibility
By including all of the components from direct sample conversion to purification, the Cells-to-CpG™ System provides a validated, flexible, simplified bisulfite conversion solution that is ideal for real-time PCR, CE, and next generation sequencing workflows. Choose a 50-reaction format or 2x96-plate format for more flexibility in performing your bisulfite conversion. Validated controls, including the Cells-to-CpG™ Bisulfite Conversion and Quantitation Control Kit as well as the Cells-to-CpG™ Methylated and Unmethylated Control Kit, are available to use through bisulfite conversion and amplification of the bisulfite-converted DNA.

Quantitatively Reliable Results The Cells-To-CpG™ Bisulfite Conversion Kit is designed to provide quantitatively reliable results verified with available controls. DNA recovery after conversion correlates linearly to the input amount with no bias observed between non-methylated and methylated samples (Figure 3).

Streamlined Bisulfite Conversion Workflow with Efficient Application
Starting with cells, tissues, blood, FFPE samples, or purified DNA, directly lyse and denature sample for subsequent bisulfite conversion, desalting, and desulfonation. The protocol takes less than 3 hours, which is a significant reduction in time compared to other available methods that require at least 18 hours.
For Research Use Only. Not for use in diagnostic procedures.

Specifications

Format: Tube(s)
Gender: Male
Species: Human
Includes: Control Primers & Template(s)
Product Size: 1 kit
Control Template: Bisulfite-Converted Genomic DNA, Non-Bisulfite-Converted Genomic DNA
Shipping Condition: Dry Ice

Contents & storage

Store kit components at –20 °C for long-term storage or at 4 °C for short-term storage.

Contains:
Unconverted Human Male gDNA (Control Sample 1) (100 ng⁄µL) 50 µL
Converted Human Male gDNA (Control Sample 2) (10 ng⁄µL) 20 µL
20 x Non-Conversion Control Primer Mix 20 µL
20 x Conversion Control Primer Mix 20 µL
20 x Quantitation Control Primer Mix 20 µL
20 x Long Amplicon Control Primer Mix 20 µL

Documents

Manuals & protocols