EZ-96 DNA Methylation-Gold Kit

D5008 / D5007


EZ-96 DNA Methylation-Gold Kit

D5008 / D5007

Cat # Name Size Price Quantity
D5008 EZ-96 DNA Methylation-Gold Kit (Deep-well) 2 x 96 Rxns $391.00
+ -
D5007 EZ-96 DNA Methylation-Gold Kit (Shallow-Well) 2 x 96 Rxns $391.00
+ -

Documents


Complete bisulfite conversion of GC-rich DNA in less than 3 hours.

Highlights


  • 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.

Description


The EZ-96 DNA Methylation-Gold Kit allows high-throughput (96-well spin-plate) conversion of unmethylated cytosines into uracil in less than 3 hours. Our optimized bisulfite chemistry combines heat denaturation and bisulfite conversion of input DNA to reduce incubation time and ensure conversion rates >99%. Recovered bisulfite-converted DNA is ideal for PCR amplification for downstream analyses including 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. 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 3 hours
Recovery >75%
Sample Source Purified genomic DNA, endonuclease-digested DNA, linearized plasmid DNA, etc. DNA should be high-quality and RNA-free.

Product FAQ


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?

Citations


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

Reviews


“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.”

- Murali B. (Centre for DNA Fingerprinting and Diagnostics)

Kit Components