mutation Posted March 19, 2007 Posted March 19, 2007 Hi everyone, I am new in Q-PCR, and using My own enzymes and dNTP mix. I do have everything what are in Kit contents but not SYBR Green buffer. I need SYBR Green buffer's recipe to start my experiment. I will be very glad if someone helps me with the recipe. thanks mutation
CharonY Posted March 19, 2007 Posted March 19, 2007 The buffer does not depend so much on the dye but rather on the polymerase that you use. Most polymerase buffer consist of a basic Tris-buffer (ph 8-8.8), Mg, and a mix of KCl and (NH4)2SO4. Other ingredients (e.g. BSA, Triton X-100, betaine etc. are possible but not essential). In short, if you use your own polymerase, just add the buffer for that enzyme.
Bluenoise Posted March 22, 2007 Posted March 22, 2007 Ditto. Just setup the reaction like any other PCR but with the addition of an appropriate amount of Sybrgreen.
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