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?
“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)
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