1 reply to this topic

[mstefani]

Hi,
I´m starting DNA methylation analysis and I have millions of questions! I´m a novice at this field and I have to construct everything from start to the end, so I hope that somebody could help me!!!
At this moment I have one problem about the "Hot Start" PCR reagents. I haven´t use hot start Taq polymerase before so what would be good one. I have found so many candidate that I don´t know what to choose. Polymerase can´t contain 3´-5´ exonuclease activity. And any extra Mg2+ will harm the result.

Thankyou, very many beforehand and I will be bag with many new questions....

-Karoliina

[pcrman]

Hi Karoliina,

Regarding hotstart taq, I encourage you to use it which may make a big difference. I use JumpStart RedTaq from Sigma and also recommend it to you.

To start your methylation experiment, use some DNA modificatin kit if you can afford it which can save you time and trouble. I have been using a kit from chemicon for a couple of years and I am satisfied with the results, although there are other options.

Good luck.