In my experiment using PCR and Gel Electrophoresis is required, since in our school only Real time PCR is available , I wounder if there would be a possibility to use Real time PCR to do my experiment.
Yes you can use real time PCR. For that you may use sybergreen technology. With this buffer you can use your existing primers. You will need to standardize the primer concentration as sybergreen binds to all dimers and by default will also bind to primer dimers. You can find out Tm of your primers and use melt curve analysis. Restricting product size between 80 to 150-200 helps.
Dear Ali
We do this regularly. You can open the tubes an load it equally like it would be a normal endpoint PCR. Success
René
Thank you all, I found it out that the Real time PCR does have both Real time and End point option and the only thing I should do is to deactivate data collection at the end point then it works as normal End point PCR.
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