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.|
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).
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.
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.
Xu, R. et al. Circulating tumour DNA methylation markers for diagnosis and prognosis of hepatocellular carcinoma. Nat Mater 2017
Pisanic, TR. et al. Long interspersed nuclear element 1 retrotransposons become deregulated during the development of ovarian cancer precursor lesions. Am J Pathol 2019
Januar, V. et al. BDNF promoter methylation and genetic variation in late-life depression. Transl Psychiatry 2015
MARIA VICTORIA GARCIA ORTIZ
INSTITUTO MAIMONIDES INVESTIGACION BIOMEDICA CORDOBA
Mar 17 2021
More effective in less time.
Quick, easy and effective bisulfite modification system. This improved version that considerably saves incubation time, is a great success for lovers of epigenetics.
Application Area: Bisulfite conversion for DNA methylation analysis
Johns Hopkins Medicine
Mar 16 2021
Reliable products with consistent results.
EZ DNA Methylation-Lightning™ Kit is the bisulfite conversion kit we have been using for years. It appeared that this is one of the kits producing pure converted DNA and would not affect downstream applications such as digital PCR which requires high purity of DNA. It is a bead-based method, which is much easier to use than others. We use it for different starting concentrations from 100 pg to 2000 ng. We get consistent results from this kit all the time.
Application Area: Bisulfite conversion of human DNA
Johns Hopkins University
Mar 16 2021
High quality methylation results.
We experience very high-quality results with this product and find the results reproducible and very reliable. We have been using Zymo methylation kits for years now, first the gold kit and now we are mainly using the lighting kit. Tech support is always very helpful and responsive whenever we have questions.
Application Area: Whole genome sequencing data
Dec 17 2020
Easy to use bisulphite conversion kit
The lightning kit is super-easy to use, efficient and reliable resource for bisulphite conversion. Great tool for methylation research.
Application Area: Methylation
Weizmann Institute of Science
Dec 05 2020
Great results, easy to use.
Easy to use, easy to scale down to single cell platforms and automation. Great results!
Application Area: Study DNA methylation during early development
|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|
EZ DNA Methylation-Lightning Kit
|D5030-E||EZ DNA Methylation-Lightning Kit (CE-IVD)||50 Rxns.|
|D5031-E||EZ DNA Methylation-Lightning Kit (CE-IVD)||200 Rxns.|
|D5030T||EZ DNA Methylation-Lightning Kit||10 Rxns.|
|D5030||EZ DNA Methylation-Lightning Kit||50 Rxns.|
|D5031||EZ DNA Methylation-Lightning Kit||200 Rxns.|