- 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.
|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?
Q2: Is an incubation with desulphonation buffer for longer than 20 minutes recommended?
Q3: Does bisulfite conversion only occur in a CpG context?
Q4: Tips for bisulfite primer design?
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?
Researchers studying circulating unmethylated and methylated preproinsulin (INS) DNA in type 1 diabetes (T1D) sought to test the differences between subjects with new-onset T1D and controls. DNA was extracted from serum using the ZR Serum DNA Kit and bisulfite converted with the EZ DNA Methylation Kit or the EZ DNA Methylation-Lightning Kit. Droplet digital PCR was used to measure unmethylated and methylated serum levels. The data demonstrated that elevations in both unmethylated and methylated INS DNA occurs in new-onset T1D and that levels of these DNA species change during T1D evolution. The use of methylation levels as potential biomarkers for diabetes may prove to be a novel tool in the near future.Fisher MM, et al. (2015) Elevations in Circulating Methylated and Unmethylated Preproinsulin DNA in New-Onset Type 1 Diabetes. Diabetes. 2015 Nov;64(11):3867-72. doi: 10.2337/db15-0430. Epub 2015 Jul 27.
The EZ DNA Methylation-Lightning Kit was used to bisulfite convert DNA from prefrontal cortex samples of patients with psychiatric diseases. The researchers then used a recently developed bisulfite padlock probe-based approach for ultra-deep mapping of modified cytosines in patients affected or unaffected by major psychosis. Their findings revealed significant epigenetic differences between patients and controls and also identified interesting epigenetic age effects.Pal M et al. (2015) High Precision DNA Modification Analysis of HCG9 in Major Psychosis. Schizophr Bull. 2015 Jun 15. pii: sbv079.
In order to investigate why endogenes are less susceptible to RNA silencing than transgenes, researchers from Germany used the EZ DNA Methylation-Lightning Kit to bisulfite convert DNA from locally and systemically silenced transgenic plant leaves. After amplifying the bisulfite-converted DNA using ZymoTaq polymerase and bisulfite sequencing, the researchers were able to conclude that cDNA transgenes are prone to post-transcriptional gene silencing and RNA-directed DNA methylation while endogene-resembling transgenes are resistant to systemic silencing and DNA methylation.Dadami E. et al. (2014) An endogene-resembling transgene is resistant to DNA methylation and systemic silencing. RNA Biology. 11:7, 1-8
“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.”