I looking controls for check the methylation conversion in my samples. I know there are full methylated and unmethylated DNA. How is the strategy for measure the conversion?
I usually test the conversion through PCR. I use a Zymo Research kit. Basically there is a converted pattern and another does not, it uses the same primers and it checks whether or not the sample is converted.
maybe these links can help you:
- https://www.zymoresearch.com/pages/bisulfite-beginner-guide#converted%20dna%20quantification
- https://epigenie.com/keeping-dna-methylation-experiments-in-check/
- Article Performance Evaluation of Kits for Bisulfite-Conversion of D...
Goof luck
Hi,
The bisulfite conversion can be checked using Methylation Specific-PCR or MS-PCR, Bisulfite sequencing and also Methylation based microarrays.
If your kit is BisulFLASH DNA Modification Kit which works better in real time MSP and its quicker or if that's Methylamp DNA Modification Kit is designed to be compatible with Illumina workflows and next-generation sequencing.The conversion also depends on downstream application as there will be variation in the fragment sizes.
Yes as above researcher mentioned Zymo Research has guide for beginners
https://www.zymoresearch.com/pages/bisulfite-beginner-guide which would be greatly helpful.
Also try this articles below
Article DNA Methylation: Bisulphite Modification and Analysis
https://www.labome.com/method/DNA-Methylation.html
Article DNA Methylation Analysis: Choosing the Right Method
Thanks, Danielle Bruno when you say DNA patterns you talk about lambda DNA (negative control) and HeLa Genomic DNA (positive control)?
Thanks for the help.
Alvaro Fernando Nieto, Just beware for a negative control, most cell lines are immortalised and have atypical methylation profiles so may not be a great negative control.
However the Zymo kits work great for conversion.
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