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Well adding Sybr green in a PCR mix


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4 replies to this topic

#1 caroGM

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Posted 26 August 2004 - 01:09 AM

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

:) :D

#2 RealTime

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Posted 26 August 2004 - 03:56 AM

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.

#3 RealTime

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Posted 26 August 2004 - 04:16 AM

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

good luck.

#4 caroGM

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Posted 26 August 2004 - 04:29 AM

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

#5 RealTime

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Posted 26 August 2004 - 04:41 AM

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

Cheers




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