EZ-96 DNA Methylation-Gold Kit
D5008 / D5007
- High-throughput (96-well) bisulfite conversion of DNA in less than 3 hours. A coupled heat denaturation/conversion reaction step streamlines the conversion of non-methylated cytosines into uracil.
- Desulphonation and recovery of bisulfite-treated DNA with a 96-well plate.
- Recovered DNA is ideal for downstream analyses such as PCR, endonuclease digestion, sequencing, microarrays, etc.
|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.|
|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||3 hours|
|Sample Source||Purified genomic DNA, endonuclease-digested DNA, linearized plasmid DNA, etc. DNA should be high-quality and RNA-free.|
Q1: Tips for bisulfite primer design?
Q2: Is an 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?
The authors used the EZ-96 DNA Methylation-Gold Kit to bisulfite convert lymphocyte DNA and subsequently analyzed the bisulfite-treated DNA with the Illumina Infinium HumanMethylation450 BeadChip Array (450k array). In this study, the authors evaluated the quality and characteristics of the probes used in the 450k array, especially with respect to detection of sex-specific differences in DNA methylation levels.Chen YA, Lemire M, Choufani S, Butcher DT, Grafodatskaya D, Zanke BW, Gallinger S, Hudson TJ, Weksberg R. (2013) Discovery of cross-reactive probes and polymorphic CpGs in the Illumina Infinium HumanMethylation450 microarray. Epigenetics. 8(2):203-9
“This protocol allows for bisulfite conversion in about 3 hours. This fast conversion is particularly relevant to our experiments, which allows us to PCR amplify and clone the products in 1 day, saving time without loss of quality.”
- Joseph M. (Sanford-Burnham Medical Research Institute)
“It makes gene specific DNA methylation analysis very simple and anyone with modest technical background can easily perform the experiment using this kit.”