McClean: EDTA (0.5M)
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Overview
EDTA is a synthetic amino acid and an agent that contains a typically organic ligand that bonds to a central metal atom. Enzymes that synthesize or modify nucleic acids (like polymerases, ligases, kinases, or nucleases) depend on Mg2+. With EDTA, it can stop the reactions between the enzymes and nucleic acids. EDTA is also in many buffers that store DNA (like TE buffer) removes the metal cofactors.
Materials
- 186.1g disodium ethylenediamine tetraacetate •2H2 (Na2EDTA)
- 800mL of dH2O
- 50mL NaOH
- Aliquots
Procedure
- Step 1 - Measure out 186.1g of Na2EDTA into a tall graduated cylinder
- Step 2 - Add in 800mL of dH2O and stir vigorously on a magnetic stirrer with a stir bar
- Step 3 - Using glass electrode (?), identify the pH of the solution
- Step 4 - If the pH is not around 8.0, carefully add in NaOH to adjust the pH
- Step 5 - Bring the solution up to 1L and stir
- Step 6 - Dispense the solution into aliquots
- Step 7 - Sterilize by autoclaving
Other Information
- If the pH of the solution is not adjusted to 8.0, EDTA will not dissolve.
- Store at room temperature
Notes
Please feel free to post comments, questions, or improvements to this protocol. Happy to have your input!
- List troubleshooting tips here.
- You can also link to FAQs/tips provided by other sources such as the manufacturer or other websites.
- Anecdotal observations that might be of use to others can also be posted here.
'''Jacquelyn Wong''':
References
Contact
- Megan N McClean 14:01, 20 July 2011 (EDT)
or instead, discuss this protocol.