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!
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...
I will use between 100 up to 200nm final SBGI concentration.
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 ?
It would be cheaper to use free SGI solution, but u can use the 2X master mix as well.