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Cat. No.: DBCMB-96 (for 96T)

Cat. No.: DBCMB-384 (for 4×96T)

Description

DNA Bisulfite Conversion Kit with Magnetic Beads, also known as the magnetic bead-based DNA bisulfite modification kit, magnetic bead-based DNA bisulfite conversion kit, magnetic bead-based DNA methylation conversion kit, or magnetic bead-based DNA methylation kit, is a reagent kit that allows for simple, fast, efficient, and high-throughput conversion of cytosine (C) in DNA to uracil (U) using bisulfite conversion, followed by purification and recovery using magnetic beads. This kit is designed for easy conversion of non-methylated cytosine (C) to uracil (U), while 5-methylcytosine (5mC) remains unconverted. It is used for subsequent PCR or high-throughput sequencing to determine whether specific CpG sites have undergone methylation modifications.

This kit is highly suitable for high-throughput 96-well plate operations and can also be used for routine single-tube operations. For the convenience of 96-well plate operations, the kit includes a 96-well deep-well plate.

DNA methylation is an important epigenetic modification that involves the covalent binding of a methyl group to the fifth carbon atom of cytosine in genomic DNA, resulting in 5-methylcytosine (5mC). In eukaryotic genomes, 5-methylcytosine is the most common chemically modified base, and CpG dinucleotides are the primary methylation sites in the genome.

DNA methylation plays a significant role in regulating gene expression, maintaining chromatin structure, genomic imprinting, X-chromosome inactivation, embryonic development, and other biological processes. It has profound effects on aging and the development and progression of diseases. DNA methylation has been closely linked to various diseases, such as cancer, mental disorders, metabolic diseases, autoimmune diseases, and more. DNA methylation can serve as a molecular biomarker or target for precise diagnosis and treatment of diseases.

Bisulfite conversion is considered the gold standard for DNA methylation analysis due to its high conversion rate (>99%) and high reproducibility. In this method, DNA is treated with bisulfite, and non-methylated cytosine (C) undergoes a series of sulfonation, deamination, desalination, and desulfonation reactions, ultimately converting non-methylated cytosine (C) into uracil (U), while methylated cytosine (C) remains unchanged.

Components

• Conversion Solution Powder
• Solution I (Protective Solution)
• Solution II (Reaction Solution)
• Solution III (Wash Buffer, add 160 ml of anhydrous ethanol before first use)
• Solution IV (Elution Buffer)
• Magnetic Beads

Features

• This reagent kit combines thermal denaturation with bisulfite conversion to process DNA samples. Subsequently, magnetic beads are used to purify the samples, and the desalination and desulfonation reactions are integrated into the magnetic bead purification process. This streamlines the experimental procedure, eliminating the need for centrifugation and simplifying the operation. It is suitable for automated high-throughput platforms and can be completed within 3 hours.
• The methylated DNA processed by this reagent kit can be used for various downstream methylation analyses, such as Methylation-specific PCR (MSP), Bisulfite sequencing PCR (BSP) for amplified products after bisulfite modification, high-throughput sequencing, restriction enzyme analysis, and more.
• This reagent kit offers high conversion rates and excellent recovery rates. It is suitable for DNA samples with an initial quantity ranging from 10 ng to 2 μg, with optimal results obtained within the range of 0.5 to 2 μg.

Storage

Store at room temperature, effective for 3 months. The magnetic beads should be stored at 4°C and are effective for one year; all other components are valid for at least one year when stored at room temperature.

Note

• This reagent kit utilizes magnetic beads for the final purification and recovery of DNA samples. A magnetic separator should be provided separately. If using the deep-well plate provided with the kit for processing 96 samples or more, an appropriate 96-well magnetic separation rack should be provided separately. This reagent kit can also be used for single-tube conversion and magnetic separation operations using regular centrifuge tubes.
• The conversion solution is recommended to be prepared and used immediately. Each tube of conversion solution powder can be used for up to 10 experiments after dissolving. Unused conversion solution can be stored at room temperature or 4°C for one day or at -20°C for one month.
• The magnetic bead suspension may settle after standing. Before use, be sure to vortex or invert several times to mix thoroughly.
• Users should provide their own anhydrous ethanol. Solution IV (wash buffer) should be mixed with 160 ml of anhydrous ethanol before the first use, and subsequently, ensure the cap is tightly closed and label the bottle.
• The conversion solution powder and Solution I (protective solution) may be harmful or irritating to the human body. Handle with care during operation, and use effective protective measures to prevent direct contact with the body or inhalation.
• This product is intended for scientific research purposes by professionals only and should not be used for clinical diagnosis or treatment, food, or drugs. It should not be stored in ordinary residential settings.
• For your safety and health, please wear laboratory attire and disposable gloves while handling.

Only for research and not intended for treatment of humans or animals

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