EZ DNA Methylation-Gold Kit

D5006 / D5005

EZ DNA Methylation-Gold Kit

Cat # Name Size Price Quantity
D5006 EZ DNA Methylation-Gold Kit 200 Rxns $596.40
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D5005 EZ DNA Methylation-Gold Kit 50 Rxns. $177.60
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  • Complete bisulfite conversion of DNA in less than 3 hours.
  • Desulphonation and recovery of bisulfite-treated DNA with a spin column.
  • Recovered DNA is ideal for downstream analyses such as PCR, endonuclease digestion, sequencing, microarrays, etc.

The EZ DNA Methylation-Gold Kit is a refinement of our popular EZ DNA Methylation kit and uses heat denaturation instead of chemical denaturation of the input DNA. This allows for the denaturation and bisulfite conversion steps to be consolidated into one step, leading to a much-reduced incubation time. Recovered bisulfite-converted DNA is ideal for PCR amplification for downstream analyses including 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.
Conversion >99%
Elution Volume ≥ 10 µl
Equipment Microcentrifuge and thermocycler with heated lid
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.
Supplemental Info

Leaving the desulphonation buffer on the column longer than recommended will cause more degradation and subsequently result in lower yields.

Bisulfite conversion will work regardless of context, so the kits are compatible with genomic DNA derived from plants and other species with high non-CpG methylation levels.

ZymoTaq DNA Polymerase has been specifically designed for use in bisulfite amplification reactions. ZymoTaq is available as a stand-alone polymerase (E2001/E2002), PreMix (E2003/E2004), and qPCR PreMix (E2054/E2055).

> 50 bp.

Following bisulfite treatment of genomic DNA, nonmethylated cytosine residues are converted into uracil. The recovered DNA is typically A, U, and T-rich. The original base-pairing no longer exists. Instead, it is single stranded with limited non-specific base-pairing at room temperature. The absorption coefficient at 260 nm resembles that of RNA. Use a value of 40 μg/ml for Ab260 = 1.0 when determining the concentration of the recovered bisulfite-treated DNA. To visualize, run converted DNA on agarose gel then chill the gel on ice for 30 minutes. The expected smear will be between 100-1500bp.

Poor conversion efficiency and low yields can be due to a variety of different experiment-specific conditions. Please contact Technical Support to discuss your specific experimental conditions and further troubleshoot with a product specialist.

Converted DNA should be eluted in M-Elution Buffer to keep the converted DNA stable for long term storage. If stored properly for long term (<-20C), the samples should last longer than a month. Minimize freeze/thawing to keep the bisulfite converted DNA stable.

“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)

“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)

Cat # Name Size Price
D5002-4 M-Wash Buffer 24 ml $30.00
D5001-4 M-Wash Buffer 6 ml $12.00
D5001-5 M-Desulphonation Buffer 10 ml $18.00
D5002-5 M-Desulphonation Buffer 40 ml $50.40
D5005-2 M-Dilution Buffer-Gold 1.5 ml $12.00
D5006-2 M-Dilution Buffer-Gold 7 ml $12.00
D5005-3 M-Binding Buffer 30 ml $24.00
D5002-6 M-Elution Buffer 4 ml $12.00
D5001-6 M-Elution Buffer 1 ml $12.00
D5006-3 M-Binding Buffer 125 ml $69.60
D5005-6 M-Dissolving Buffer 500 µl $12.00
D5006-6 M-Dissolving Buffer 1.2 ml $18.00
C1001-50 Collection Tubes 50 Pack $16.50
C1004-50 Zymo-Spin IC Columns 50 Pack $58.30
D5001-1 CT Conversion Reagent 1 tube for 10 Conversions $12.00