Preparation of NMR Buffers Protocol

3. Preparation of NMR Buffers Protocol:

3.1.Stock Solutions:

3.1.1  0.1M stock solution of Na2HPO4.

        Weigh 141.96g of Na2HPO4 (anhydrous) and dissolve to 1000 ml (one Liter)     with ddH2O.

3.1.2  0.1M stock solution of NaH2PO4.

           Weigh 119.98g of NaH2PO4 (anhydrous) and dissolve to 1000 ml (one Liter) with ddH2O.

NB: The weight of each salt needed to prepare one liter of a 1M solution varies, depending on the hydrated state of the salt.

3.1.3    .50m M stock solution of DSS.

           Weigh 1.0915g of DSS and dissolve to 100 ml with ddH2O.

3.1.4. NaOH solution.

3.1.5. HCl solution.

3.2. NMR Buffer number 1:

3.2.1. Measure 28.85 ml of the 1M stock solution of Na2HPO4 and 21.15 ml of the      1M stock solution of NaH2PO4 into a 1000 ml graduated cylinder.

3.2.2. Mix and add ddH2O to 850 ml.

3.2.3. Add 100 ml of D2O.

3.2.4. Add 10 ml of the 50 mM DSS stock solution.

3.2.5. Mix well and adjust to pH to 7.0 (if necessary) with HCL or NaOH solution.

3.2.6. Make volume up to 1000 ml with more ddH2O

3.2.7. Store in a labeled, dated bottle in the refrigerator.

3.2.8. Check for impurities before use.

3.3.NMR Buffer number 2:

3.3.1. Measure 28.85 ml of the 1M stock solution of Na2HPO4 and 21.15 ml of the    1M stock solution of NaH2PO4 into a 1000 ml graduated cylinder.

3.3.2. Mix and add ddH2O to 980 ml.

3.3.3. Mix well and adjust to pH to 7.0 (if necessary) with HCL or NaOH solution.

3.3.4. Make volume up to 1000 ml with more ddH2O

3.3.5. Store in a labeled, dated bottle in the refrigerator.

3.3.6.  Check for impurities before use.