IGEM:Brown/2007/Lab Protocols/10X Ligation Buffer Recipe: Difference between revisions
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Adam.Emrich (talk | contribs) (New page: For 10 ml of 10X ligation buffer: Ingredients: 1. 2 mls of 1 M stock Tris at pH=7.6. 2. 102 mg of MgCl2. 3. 77 mg of DTT 4. 5 mg of 1 mg/ml BSA. 5. 28 mg of ATP. The volume is then...) |
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Revision as of 10:07, 7 June 2007
For 10 ml of 10X ligation buffer:
Ingredients: 1. 2 mls of 1 M stock Tris at pH=7.6. 2. 102 mg of MgCl2. 3. 77 mg of DTT 4. 5 mg of 1 mg/ml BSA. 5. 28 mg of ATP.
The volume is then brought up to 10 mL, and the pH is checked.
The resultant buffer has the following concentrations: 1. 0.2 M Tris pH=7.6. 2. 50 mM MgCl2. 3. 50 mM DTT. 4. 500 ug/ml BSA. 5. 5 mM ATP.
1ml aliquots are stored at -80 C for long term storage. 50 ul aliquots are stored at -20 C for short term storage. Freeze thaw at most 2-3 times, then discard.
