Bisulfite Conversion Kit for DNA Methylation Analysis (Magnetic Beads – based)

Description

Magnetic Beads – based Bisulfite DNA Conversion Kit from BioDynami

Highest Conversion Efficiency, Purity, and Concentration of converted DNA

• Conversion efficiency >99.5 %
• Low elution volume (20-50 µl) without yield loss
• Extremely high purity: Compatible with PCR, qPCR, methylation-specific PCR (MS-PCR), NGS, Pyrosequencing, COBRA, and Methylation Arrays
• Input DNA range: 1-1000 ng
• Bead-based:  Fast, easy, automation friendly – no centrifuge, no column, no vacuum

The BioDynami DNA Bisulfite Conversion Kit (Magnetic Beads) was developed for the detection of genomic DNA methylation at single base resolution at high efficiency and at ease.

Genomic DNA is denatured to single-stranded DNA and treated with sodium bisulfite in one single procedure. Sodium bisulfite selectively converts unmethylated cytosines into uracils through deamination, while the methylated cytosines (including both 5-methylcytosine and 5-hydroxymethylcytosine) are not changed.

The conversion is typically followed by PCR, in which uracils are further converted to thymines. As a result, all unmethylated cytosines are converted into thymines, and all methylated cytosines remain cytosines. The conversion difference of methylated and unmethylated cytosines can be detected and identified by comparison to the original genome.

Downstream applications for analyzing DNA methylation by bisulfite-conversion include PCR, qPCR, methylation-specific PCR (MS-PCR), COBRA, pyrosequencing, Methyl-seq by Next Generation Sequencing (NGS), and methylation-based arrays, all of them requiring high converted DNA purity and concentration.

BioDynami have developed an optimized protocol applying engineered magnetic beads coated with carboxyl-groups and reversibly binding to the converted DNA, as well as specially designed buffers, for desulphonation and purification of the converted DNA at highest recovery rates (>70%), and in smallest elution volumes, to obtain extremely pure (OD 260/A280 ≥1.8; OD 260/A230 ≥1.5) and concentrated converted DNA (elution volumes are 20-50 ul), enabling all applications.

Data Examples

Conversion PCR Data

Conversion qPCR data

Kit Contents (48 rxns,  1-1000 ng  gDNA input  per rxn):

Conversion Buffer:  6 x 1.2 ml

Conversion Beads: 15 ml

Desulphonation Buffer: 5 ml

Elution Buffer: 3 ml