EZ-96 DNA Methylation-Lightning Kit
D5032 / D5033
- Fastest 96-well bisulfite conversion kit for complete, high-throughput (96-well) 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||Deep-well: ≥ 15 µl
Shallow-well: ≥ 30 µl
|Equipment||Thermocycler with heated lid, swinging-bucket centrifuge with plate carriers.|
|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: Tips for bisulfite primer design?
Q2: Which polymerase is recommended for amplification from bisulfite converted DNA?
Q3: Does bisulfite conversion only occur in a CpG context?
Q4: Is an incubation with desulphonation buffer for longer than 20 minutes recommended?
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