EZ-96 DNA Methylation Kit

D5004 / D5003


EZ-96 DNA Methylation Kit

D5004 / D5003

Cat # Name Size Price Quantity
D5004 EZ-96 DNA Methylation Kit (Deep-Well) 2 x 96 Rxns $380.00
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D5003 EZ-96 DNA Methylation Kit (Shallow-Well) 2 x 96 Rxns $380.00
+ -

Documents


Streamlined bisulfite treatment of DNA.

Highlights


  • Streamlined, proven procedure for bisulfite conversion of DNA.
  • Desulphonation and recovery of bisulfite-treated DNA with a 96-well spin-column plate.
  • Recovered DNA is ideal for downstream analyses including PCR, endonuclease digestion, sequencing, microarrays, etc.

Description


The EZ-96 DNA Methylation Kits features a high-throughput (96-well spin-plate), simplified procedure that streamlines bisulfite conversion of DNA. Cytosines undergo a three-step reaction with sodium bisulfite during which the cytosine is converted into uracil. The innovative in-column desulphonation technology eliminate otherwise cumbersome precipitations. The kit is designed to reduce template degradation and minimize DNA loss during treatment and clean-up, while ensuring complete conversion of the DNA. Purified, converted DNA is ideal for downstream analyses including PCR amplification, endonuclease digestion, sequencing, microarrays, etc. Note: Catalog # D5004 is recommended for use with the Illumina Infinium MethylationEPIC BeadChip array.

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. Catalog # D5004 is recommended for use with Illumina Infinium MethylationEPIC BeadChip array.
Conversion >99%
Elution Volume ≥ 15 µl for Deep-well
≥ 30 µl for Shallow-well
Equipment Thermocycler with heated lid, swinging-bucket centrifuge with plate carriers.
Input 500 pg - 2 µg of DNA
Processing Time 12-16 hours
Recovery >80%
Sample Source Purified genomic DNA, endonuclease-digested DNA, linearized plasmid DNA, etc. DNA should be high-quality and RNA-free.

Product FAQ


Q1: What leads to poor conversion efficiency/ low yields?

Q2: How to quantify / visualize converted DNA?

Q3: What is the minimum DNA size that can be recovered?

Q4: How long is bisulfite converted DNA stable at -20 °C?

Q5: Does bisulfite conversion only occur in a CpG context?

Q6: Is an incubation with desulphonation buffer for longer than 20 minutes recommended?

Q7: Which polymerase is recommended for amplification from bisulfite converted DNA?

Q8: Tips for bisulfite primer design?

Citations


Researchers from Barcelona recently used the EZ-96 DNA Methylation Kit to successfully validate the new MethylationEPIC BeadChIP (Infinium 850K array, Illumina) with FFPE samples. The results validate the 850K array as a highly reliable genomic platform for the study of DNA methylation patterns and levels in the human genome. The data also represents an extremely valuable tool to decipher how DNA methylation changes in unexplored territories, such as enhancer sequences, contribute to cell homeostasis and human diseases.

Moran S, et. al. (2015) Validation of a DNA methylation microarray for 850,000 CpG sites of the human genome enriched in enhancer sequences. Epigenomics. 2015 Dec 17.

In a genome-wide study investigating DNA methylation levels in acute myeloid leukemia cells, researchers used the EZ DNA MethylationTM Kit to bisulfite convert genomic DNA obtained from clinical samples. PCR amplification followed by bisulfite sequencing of the converted DNA helped to show that patients diagnosed with acute myeloid leukemia have distinct methylomes in their cancer cells compared to normal cells.

Saied M, et. al. (2012) Genome wide analysis of acute myeloid leukemia reveal leukemia specific methylome and subtype specific hypomethylation of repeats. PLOS. 2012 Mar 29.

Kit Components