Bisulfite Sequencing - detect DNA Methylation

Hey Friends,
todays topic will be: Bisulfite Sequencing, which is a method for the detection of DNA Methylation.
Hope you enjoy it.
Cheers,
Henrik

[@henrikslab]

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[@aamaramuzaffar3565]

You way of teaching is amazing

[@NickoReese]

Great video! Thank you for explaining it so fully.

[@saleemamehboob7549]

Great explanation. Very comprehensive summary with good animation👍👍👍

[@xXArtimixXx]

Beautifully and quickly explained. Thanks for the video, you earned a sub!

[@nanashalev7612]

Amazing explanation, right on the point. Thanks :)

[@TheSummerOf86]

Great video, short and simple, nice graphics. Thank you!

[@cankale8002]

I really like these kinds of videos where you understand every part of the topic. It was really helpfull.

[@elanafarrell5929]

This is super informative and easy to understand thank you

[@quinattasneemrafique536]

Amazing crisp explanation. Thanks a lot

[@nivethanimmy9985]

Explanation was clear. thank you.

[@orxanibayev340]

Helped a lot, many thanks!

[@mariociencia12]

I just donated about 2 dollars for the video. But the video deserve much more than that

[@Mohannad23801]

This was very useful thanks a lot!

[@bartkuk1]

Very clear explanation. Thank you 😊

[@navyanandhanaofficial]

Very nice video thanks for sharing

[@nabyl740]

Nice channel and nice videos 👌

[@Daisy-lg7sq]

Thank you so much for clear explanation ❤

[@genicadelara5243]

Thank u so much. This is helpful.

[@ebtihal1990]

Great explanation, thank you for your help

[@curioussepia7677]

Thank you ,it was helpful.

[@naifalharbi6730]

Amaaaaaazing thank u

[@Displ4c]

Awesome 🙏🏻 thank you

[@bhavikatiwari2746]

Thanks for this easy explanation!

[@henrikslab]

Thanks for watching!

[@hadijohni23]

so grateful sir

[@younesabuelayyan4520]

well explained, thanks.

[@mdkhurshid7598]

Excellent

[@c.wredacted2589]

Thank you so very much

[@sadafyasmeen3111]

Well, how are we gonna use the same primers for the template whose Cs are changed to Ts in PCR?

[@omercetin1683]

Good exp.

[@rougang_0471]

I'm wondering how to confirm the specific T(which is the unmethylated cytosine) after PCR? And also thanks for your video which is clear and easy to understand!

[@henrikslab]

You wonder how distinguish between the “new“ T (unmethylated cytosine) and the “normal“ T in the DNA, correct? This works only by comparing the untreated DNA with bisulfite treated DNA... (sequencing) e.g. Untreated DNA A T C A G G Treated DNA A T T A G G the first T was originally also a T the second T was initially an unmethylated C

[@dinushiarambegedara9906]

@@henrikslab Do you know a way to compare the bisulfite converted sequence with a non-converted sequence? A software maybe?

[@jinglin4358]

I know this question is a few years old but it’s possible to sequence whole genome with low concentrations of DNA ie extracted and not amplified DNA with next gen sequencing. So you can compare the sequence prior to bisulfite conversion and after.

[@yaxiu3799]

Good.

[@mayconmarcao4554]

Well done explanation! But I have to confess that the "arrows" you showed ( C > C or C > T) looks like "greater than". 3:34

[@henrikslab]

I agree! This is a bit misleading..

[@ashukhan-jr2kl]

My first like and first comment

[@henrikslab]

What an honor!

[@mariociencia12]

Valeu!

[@henrikslab]

Not again... stop it, Mario! Thank you, man!

[@markrudolf7567]

How do you know a given base was an un-methylated C and not a T? Do you to compare it the original sequence somehow?

[@jinglin4358]

Yes. You compare post PCR sequence to pre. it’s possible to sequence whole genome with low concentrations of DNA ie extracted and not amplified DNA with next gen sequencing. So you can compare the sequence prior to bisulfite conversion and after.

[@alxdr9451]

love for that still

[@mariociencia12]

The video is very nice. However, I had not understood at first glance why U is converted into T in PCR. In another video, I saw that a complementary sequence is generated in a first-round, where U is paired with A. Then, in a second-round, the primary sequence is generated, but with T paired with A. It is equivalent to the conversion of U into T. I had confused because, in the cell, the U in DNA appears to be corrected into C. If the video showed the conversion of U into T in PCR it will be perfect!

[@henrikslab]

I agree! I decided to skip the step you mentioned ´cause I put the focus on the bisulfite conversion itself. But you described it correctly! I said "certain rounds of PCR" to simplify everything, but U is paired with A and this newly synthesized strand is paired with T then! True

[@alerookoh8748]

I thought methylation mostly occurs at CpG islands alone?

[@henrikslab]

A CpG island is a region in the DNA with a high C G dinucleotide frequency and density. They are often located in promoter regions. Indeed, here we find an important part of methylation. You probably talk about CpG *sites*? Mainly methylation occurs at CpG sites, true. However, there is also methylation at CpA, CpT and CpG sites. See -> www.ncbi.nlm.nih.gov/pmc/articles/PMC5485512/

[@alerookoh8748]

Thank you for clarifying my question

[@Jojochecker123]

My last like and last comment!

[@henrikslab]

Der Kritiker Ehre