Melting Curve Analysis for semiquantitative measurement of promoter methylation - (Feb/19/2007 )

Hi, a technical question, again.

Has anybody experience in using the melting curve approach for qualitative or semiquantitative analysis of gene promoters? I have seen some older papers (Pubmed ID: 12095269) and a recent technical note in biotechniques (ID: 16989087) and very recently in Nucl Acids (17289753). I think it sounds promising for high throughput analysis of large sample sizes, but would be interested in your opinion on that! Has anybody tried it?

Regards,

Krümel

hi Kruel,

I have heard good things about it and there are qPCR machines that now have a high resolution melting curve capability for such an assay.

I have had a chat with the author of the biotechniques paper, he is now based in Melbourne, Australia in Alexander Dobrovic's lab.. .

Certainly a much cheaper, high throughput method for methylation analysis.

Although have you ever considered Sequenom?

N

Ooops, I never even heard of that. Please, tell me more about Sequenom.

Krümel


... okay been to their homepage - quite interesting, especially given the fact that we are currently working on a similar method employing MALDI-TOF based analysis (nevertheless, we thougth of using bisulfite modification insted of enzymatic cleavage). But do you have any idea, what they want for that?

Krumel,

look up the mass array epityper part of sequenom, does bisulfite analysis.

We are hoping to get one soon and are raising funds, it's about $250K AUD for the system.

Enzymatic cleavage is for SNP analysis, but it's epityper package is very attractive.

I know sequenom themselves offer it as a service in San Diego as we have sent samples to them directly, A service is being set up here in australia also.

Allright, I was just confused by the enzymatic cleavage they use to produce different sized fragments, but that is needed for the MALDI analysis. Fingers crossed for your fund raising!
and I need to talk to my boss ... maybe he can rob a bank?

Krümel

well we may have to rob a bank also seems like politics are at play where i am.