I understand that there are differences in ion preference as far as chelation is concerned when comparing EGTA vs EDTA.
But is it ok to replace EGTA in my lysis buffer with EDTA since EDTA appears to be much more readily available and cheaper as well?
Hello Ralph Lim
No, you cannot replace EGTA with EDTA in the lysis buffer for nuclei acid extraction.
DNase I has optimal activity in buffer containing Mg2+ and Ca2+ ions. But DNase I is not active in buffers containing Mg2+ and lacking Ca2+. This indicates that Ca2+ can act as a potent enzyme activator in the presence of Mg2+. Ca2+ is known to bind tightly to DNase I and stabilize its active conformation.
As you know that compared to EDTA, EGTA has a higher affinity for calcium ions but a lower affinity for magnesium ions. Therefore, you will find EGTA in lysis buffer rather than EDTA as EGTA can efficiently remove Ca2+ and inhibit DNase I activity thereby preventing DNA degradation.
So, you will have to include EGTA in lysis buffer instead of EDTA because EGTA has a specific role to play.
Best.
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