EZ DNA Methylation-Gold Kit

D5006 / D5005


EZ DNA Methylation-Gold Kit

D5006 / D5005

Cat # Name Size Price Quantity
D5006 EZ DNA Methylation-Gold Kit 200 Rxns $497.00
+ -
D5005 EZ DNA Methylation-Gold Kit 50 Rxns. $148.00
+ -

Documents


EZ DNA Methylation-Gold Kit

Highlights


  • 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.

Description


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.

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.
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.

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


Researchers from the National Institute of Plant Genome Research in New Delhi used the EZ DNA Methylation-Gold Kit to identify the level of cytosine methylation in genomic DNA from Tomato leaf curl New Delhi virus. Through DNA methylation-specific RNA silencing, the researchers aimed to target the viral genomic regions of this virus to potentially determine an alternate defense mechanism for generating transgenic plants to prevent yield loss.

Sahu P et al. Post-transcriptional and Epigenetic Arms of RNA Silencing: A Defense Machinery of Naturally Tolerant Tomato Plant Against Tomato Leaf Curl New Delhi Virus. Plant Mol Biol Rep (2014) 32:1015-1029

The EZ DNA Methylation-Gold Kit was used to bisulfite convert DNA extracted from Thy-1 (+) and Thy-1 (-) lung fibroblasts and tissue. Data from methylation-specific PCR and bisulfite sequencing enabled researchers to show that epigenetic regulation of Thy-1 could be a novel and potentially reversible mechanism in fibrosis that may offer the possibility of new therapeutic options.

Sanders Y et al. Thy-1 Promoter Hypermethylation A Novel Epigenetic Pathogenic Mechanism in Pulmonary Fibrosis. Am J Respir Cell Mol Biol. 2008 Nov; 39(5):610-8

Researchers from China analyzed the role of DNA methylation in the life cycle of a parasitic nematode, Trichinella spiralis and characterized the methylome of three life-cycles states in this organism. Genomic DNA of T. spiralis muscle larvae, adults, and new-born larvae was isolated and methylated cytosine levels were determined by MethylC-seq.

Gao F et al. (2012) Differential DNA methylation in discrete developmental stages of the parasitic nematode Trichinella spiralis. Genome Biol. 13(10):R100.

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Reviews


“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)

Kit Components