Anyone know of a buffer that will work adequately for PCR with Takara's Primestar GXL? I have tons of enzyme but am out of buffer. Takara will sell a pack of extra buffer tubes, but it won't arrive for a month!
I have old tubes of Pfu, Taq and Vent buffers or could make up a generic buffer if I knew the pH, ionic strength and Mg2+ concentrations that GXL likes, but Takara won't say...
you can attempt to create a generic PCR buffer with common components and optimize your PCR conditions. Here's a basic recipe for a generic PCR buffer. Generic PCR Buffer:
10x Tris-HCl buffer (pH 8.3 to 9.0)
50 mM KCl
1.5 mM MgCl2 (you may need to optimize this concentration)
You can adjust the pH using concentrated Tris base or HCl. The pH range is broad because different enzymes have varying pH optima. For Taq polymerase, a pH around 8.3 is often used, but some enzymes may work better at a slightly higher pH.
Regarding Mg2+ concentration, you'll need to optimize it. Different polymerases have varying requirements for Mg2+. For a start, you can prepare a range of concentrations (e.g., 1.5 mM, 2.0 mM, 2.5 mM) and test them in your PCR reactions to see which gives you the best results.
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