Well adding Sybr green in a PCR mix - to detect quicker than by electrophoresis PCR prod (Aug/26/2004 )

I'd like to know if someone as already tried to add Sybr Green I in a PCR mix, and in which conditions (concentration), to detect PCR products with a fluorimeter or a real time thermocycleur. This is in order to gain time and not to do an electrophoresis after this end point PCR.

thank you for advance to help me!


CaroGM

Hi Caro,

U can try to add SYBR GreenI into your end point PCR (follow ie:sigma recommandations). in this case u can performe a melting curve on a real time PCR system to indentify them... but no possible quantification...

Bye.

I will use between 100 up to 200nm final SBGI concentration.

good luck.

thank you very much for your recommendations!
it's likely what i was thinking about


don't you think i should use SGI at a concentration of 1x final like in normal quantitation ?

Caro

It would be cheaper to use free SGI solution, but u can use the 2X master mix as well.

Cheers