Hello everybody,
Accidentally we have a large amount of Sigma-Aldrich DNA Ligase from T4-infected Escherichia coli and Promega T4 DNA ligase buffer.
The "correct" Sigma-Aldrich buffer is made by 500 mM Tris-HCl, 100 mM MgCl2, 50 mM DTT and 10 mM ATP.
Our Stock Promega buffer is made with 300mM Tris-HCl , 100mM MgCl2, 100mM DTT and 10mM ATP.
Can we use the Promega buffer with the Sigma-Aldrich enzyme? Does somebody did something similar?
Thank you in advance, best
Umberto
Carlos Pérez Arques
I didn't try that particular combination, but I have used ThermoScientific T4 ligase with some restriction enzyme buffer supplemented with ATP (which you don't even need because ATP is already in your Promega buffer) and it worked for me. I don't think it should be a problem.
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Laurence Stuart Dawkins-Hall
- In short your buffer will be fine
- Your Tris Molarity is near enough and crucially your Mag and NTP concentrations are identical
- Go ahead therefore and use: It shyould ork with exactly the same efficiency
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