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EZ DNA Methylation-Lightning Kit
D5030-E / D5031-E / D5030T / D5030 / D5031
- Fastest bisulfite conversion kit for complete bisulfite conversion of DNA for methylation analysis.
- Ready-to-use conversion reagent is added directly to DNA.
- High-yield, converted DNA is ideal for PCR, Methylation Specific PCR (MSP), arrays, library preps, Next-Generation sequencing, etc.
The EZ DNA Methylation-Lightning Kit can rapidly bisulfite convert and purify DNA in less than 1.5 hours. A ready-to-use conversion reagent streamlines this process – simply add the reagent directly to the sample and incubate. Desulphonation and clean-up of the converted DNA is performed on a spin-column, allowing an elution volume as low as 10 µl. The kit achieves high conversion efficiency of unmodified cytosines into uracil, ensuring accurate downstream methylation analysis.
|Applications||Purified, converted DNA is of high-quality and well-suited for downstream processes, including library preparation for Next-Generation sequencing, PCR amplification, etc.|
|Elution Volume||≥ 10 µl|
|Equipment||Thermocycler with heated lid and microcentrifuge.|
|Input||100 pg - 2 µg of DNA.|
|Processing Time||1.5 hours|
|Sample Source||Purified genomic DNA, endonuclease-digested DNA, linearized plasmid DNA, etc. DNA should be high-quality and RNA-free.|
Q1: Which polymerase is recommended for amplification from bisulfite converted DNA?
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).
Q2: Is an incubation with desulphonation buffer for longer than 20 minutes recommended?
Leaving the desulphonation buffer on the column longer than recommended will cause more degradation and subsequently result in lower yields.
Q3: Does bisulfite conversion only occur in a CpG context?
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.
Q4: Tips for bisulfite primer design?
Q5: What is the minimum DNA size that can be recovered?
> 50 bp.
Q6: How to quantify converted DNA?
For best results, keep the method of quantification consistent before and after bisulfite treatment:
- If quantifying with a NanoDrop, use dsDNA settings (50 μg/ml for Ab260 = 1.0) before treatment and use RNA settings (40 μg/ml for Ab260 = 1.0) after treatment.
- If quantifying with Qubit, use a dsDNA assay before treatment and use a ssDNA assay after treatment.
Q7: How to visualize converted DNA?
Following bisulfite treatment, DNA will be single stranded with limited non-specific base pairing at room temperature. To visualize, run the converted DNA on an agarose gel then chill the gel on ice or in an ice bath for 30 minutes. This will force enough base-pairing to allow intercalation of the ethidium bromide for the DNA to be visible. If using a Bioanalyzer or TapeStation instrument, use RNA kits and reagents to visualize the converted DNA.
Q8: What leads to poor conversion efficiency/ low yields?
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.
Q9: How long is bisulfite converted DNA stable at -20 °C?
Converted DNA eluted in M-Elution Buffer can generally be stored at -20°C for 1-3 months. If longer term storage is necessary, we recommend storing at or below -70°C if possible. Bisulfite converted DNA is less stable than dsDNA; for best results, minimize freeze-thawing of converted DNA and use as soon as possible for downstream analysis.
“Results were as good as with the formerly used EZ DNA Methylation Gold Kit but significantly reduced total time for analyses. More flexibility due to the possibility of up to 20h storage of the samples after bisulfite conversion before the cleaning procedure especially for part-time employees.”
- Sabrina S. (UK-Aachen)
“The conversion reagent no longer has to be made up from individual components and dissolved - it is supplied ready-made. The conversion reagent can be stored at room temperature - there is no need to use up all 10 uses in one month. This makes it more flexible, and avoids wastage. Shorter protocol saves time.”
- Jeremy B. (AgResearch)
|C1004-50||Zymo-Spin IC Columns||50 Pack|
|C1001-50||Collection Tubes||50 Pack|
|D5002-4||M-Wash Buffer||24 ml|
|D5001-4||M-Wash Buffer||6 ml|
|D5005-3||M-Binding Buffer||30 ml|
|D5002-6||M-Elution Buffer||4 ml|
|D5001-6||M-Elution Buffer||1 ml|
|D5006-3||M-Binding Buffer||125 ml|
|D5030-5||L-Desulphonation Buffer||10 ml|
|D5031-5||L-Desulphonation Buffer||40 ml|
|D5030-1||Lightning Conversion Reagent||1.5 ml|