Monday, January 3, 2011
1. NMR tube containing sample should be held in a plastic spinner (use the blue color spinner in 500 and 600 MHz spectrometers; use the white color spinner in 700 MHz spectrometer). Hold the sample by the top, place sample tube in the spinner and the spinner in the sample depth gauge. Push or pull the sample tube so that the depth of the sample above and below the center line of the sample depth gauge is equal. However, never exceed the lower limit (position of the adjustable white platform which should be at the line marked 5 mm 15mm) as this can damage the probe as well as the sample, as the sample will rest on the tip of the tube, not the spinner. This is important for all probes, but especially the cryoprobes installed on the 600 and 700 MHz spectrometers.
2. Remove the black cap from the top of the magnet bore. Next, press the LIFT button in upper left portion of the BSMS keyboard. Wait for the airflow (hissing sound that can be heard). Then remove the depth gauge before inserting the sample and spinner into the magnet. Pressing the LIFT button will toggle the air flow off and will drop the sample tube gently to the magnet bore where it will be positioned at the top of the probe.
3. Preparation of NMR Buffers Protocol:
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.