IGEM:Brown/2007/Lab Protocols/10X Ligation Buffer Recipe

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.