EZ-96 DNA Methylation-Direct MagPrep

D5045 / D5044


EZ-96 DNA Methylation-Direct MagPrep

D5045 / D5044

Cat # Name Size Price Quantity
D5045 EZ-96 DNA Methylation-Direct MagPrep 8 x 96 Rxns. $1,126.00
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D5044 EZ-96 DNA Methylation-Direct MagPrep 4 x 96 Rxns. $696.00
+ -

Documents


A complete and reliable bisulfite conversion kit that eliminates cumbersome DNA precipitation steps and allows DNA bisulfite conversion directly from blood, tissue, and cells without prior DNA purification.

Highlights


  • High throughput, complete bisulfite conversion of DNA directly from blood, tissue, cells, FFPE and LCM-derived samples
  • Compatible with small sample inputs – as few as 10 cells or 50 pg of DNA.
  • High throughput (96-well), automated desulphonation and recovery of bisulfite-treated DNA.

Description


The EZ-96 DNA Methylation-Direct MagPrep is a magnetic bead-based bisulfite conversion kit that allows simple and reliable DNA bisulfite conversion directly from blood, tissue, and cells without the prerequisite for DNA purification. The increased sensitivity of this magnetic bead-based bisulfite conversion kit makes it possible to amplify bisulfite-converted DNA from as few as 10 cells or 50 pg DNA. These innovations have been coupled to a magnetic bead-based clean-up for high-throughput methylation analysis. Desulphonation and clean-up of the converted DNA is performed while bound to the MagBinding Beads. This magnetic bead based bisulfite conversion kit has been designed to minimize template degradation, loss of DNA during treatment and clean-up, and to provide complete conversion of unmethylated cytosines. Recovered DNA is ideal for downstream analyses, including PCR amplification, endonuclease digestion, sequencing, microarrays, etc.

Technical Specifications


Applications Purified, converted DNA is of high-quality and well-suited for downstream processes, including library preparation for Next-Generation sequencing, PCR amplification, etc.
Conversion > 99.5%
Elution Volume ≥ 25 µl
Equipment Thermocycler with heated lid, heating element for 96-well plate, magnetic stand.
Input Samples containing between 50 pg to 2 µg of DNA. For optimal results, the amount of input DNA should be from 200 to 500 ng. The number of cells per standard treatment can range from 10-105 cells.
Processing Time 4 hours
Recovery > 80%
Sample Source Blood, tissue, cells, FFPE, LCM-derived samples, purified genomic DNA, endonuclease-digested DNA, linearized plasmid DNA, etc.

Product FAQ


Q1: Tips for bisulfite primer design?

Q2: Is incubation with Desulphonation Buffer for longer than 20 minutes recommended?

Q3: Does bisulfite conversion only occur in a CpG context?

Q4: Which polymerase is recommended for amplification from bisulfite converted DNA?

Q5: What is the minimum DNA size that can be recovered?

Q6: How to quantify / visualize converted DNA?

Q7: What leads to poor conversion efficiency/ low yields?

Q8: How long is bisulfite converted DNA stable at -20 °C?

Citations


Bisulfite treatment of DNA from fibroblast cells of Aicardi-Goutières syndrome (AGS) patients was performed using the EZ DNA Methylation-Direct Kit prior to MethylC-seq. Results showed a global loss of DNA methylation from patients with TREX1, RNASEH2, and SAMHD1 mutations, indicating epigenetic abnormalities in AGS.

Lim YW, et. al. (2015). Genome-wide DNA hypomethylation and RNA:DNA hybrid accumulation in Aicardi-Goutières syndrome. Elife.

In this publication, researchers investigated the regulatory effects of BRCA and glucocorticoid receptor (GR). Genomic DNA extracted from both ovarian cancer and normal ovarian tissue was bisulfite converted using EZ DNA Methylation-Direct Kit and BRCA1 promoter methylation was analyzed. Immunochemistry, real-time PCR, regression analysis, knockdown, and overexpression experiments were also performed to examine the relationship between BRCA1 and GR expression levels, which were found to positively correlate in cancer tissues.

Fang YY et al. (2014) Glucocorticoid receptor repression mediated by BRCA1 inactivation in ovarian cancer. BMC Cancer. 14:188.

Epigenetic changes were shown to be induced in the midbrain of adult mice by social isolation from 3-6 months of age. Genomic DNA was isolated from the midbrain of the “lonely” mice and was bisulfite converted using the EZ DNA Methylation-Direct Kit. Bisulfite pyrosequencing was performed and the researchers found that social isolation of adult male C57BL/6 mice led to global DNA methylation changes in the midbrain.

Siuda D et al. (2014) Social isolation-induced epigenetic changes in midbrain of adult mice. J Physiol Pharmacol. 65(2):247-55.

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Kit Components