- Hydrogen exchange effect on T1 and T2 measurements are reduced by steady-state pulse (sspul); Kim et. al. (2013) JBNMR 55, 249. SK versions use sspul in Agilent and Bruker sequences.
- For T2 measurement of large proteins, the relaxation time per a CPMG cycle is made half.
- A new NOE build-up pulse scheme is included; Ferrage et. al. (2010) JMR 207, 294.
- Pseudo-3D acquisition in Bruker; the relaxation parameter can be arrayed via vd lists or vc lists and it is encoded as an additional dimension (F1QF).
- Interleaved 2D acquisition in Bruker; in N15 NOE measurement, F1I is used for options of 1H saturation and no saturation.
- Bruker topspin 3 introduced changes of sequences; TC (temperature compensation), shaped 1H pulses, and reducing the CPMG cycle.
| NMR Experiment | Bruker Sequence |
|---|---|
| 15N-T1 HSQC | hsqcf3T1sk Acquire pseudo-3D; F3(1H) - F2(15N) - F1(vd list) Use sspul and TC vd list = 0.01, 0.02, .... (s) Modified from hsqct1etf3gpsitc3d |
| 15N-T1 TROSY | trosyf3T1sk Acquire pseudo-3D; F3(1H) - F2(15N) - F1(vd list) Use sspul and TC vd list = 0.01, 0.02, .... (s) Modified from trt1etf3gpsi3d.2 |
| 15N-T2 HSQC | hsqcf3T2sk Acquire pseudo-3D; F3(1H) - F2(15N) - F1(vc list) vc list = 1, 2, 3.... for loops of CPMG Use sspul and TC Relaxation time = 8ms*vc Modified from hsqct2etf3gpsitc3d.2 that uses relaxation time = vc*16*(0.45ms+0.5p30) = vc*7.92ms |
| 15N-T2 TROSY | trosyf3T2sk Acquire pseudo-3D; F3(1H) - F2(15N) - F1(vc list) Use sspul and TC vc list = 1, 2, 3.... for loops of CPMG Relaxation time = 8ms*vc Modified from trt2etf3gpsitc3d.2 that uses relaxation time = vc*16*(0.45ms+0.5p30) = vc*7.92ms |
| 15N-NOE HSQC | hsqcf3NOEsk Acquire interleaved-2D; F2(1H) - F1(15N) - F1I(l0) F1I needs 2 times td1 Use d1=2 and d20=4 for NOE Use the 1H saturation scheme by Ferrage(2010) JMR, 207,294 xau calibsk for PLDB6 |
| 15N-NOE TROSY | trosyf3NOEsk Acquire interleaved-2D; F2(1H) - F1(15N) - F1I(l0) F1I needs 2 times td1 Use d1=2 and d20=4 for NOE Use the 1H saturation scheme by Ferrage(2010) JMR, 207,294 xau calibsk for PLDB6 |
| 15N-T2HE HSQC (T2 Hahn echo to check Rex) |
hsqcf3T2HEsk Acquire pseudo-3D; F3(1H) - F2(15N) - F1(vd list) Use sspul and TC vd list = 0.01, 0.02, .... (s) Relaxation time= 7.68ms*vd/0.01 xau calibsk for PLDB19 |
| 15N-T2HE TROSY (T2 Hahn echo to check Rex) |
trosyf3T2HEsk Acquire pseudo-3D; F3(1H) - F2(15N) - F1(vd list) Use sspul and TC vd list = 0.01, 0.02, .... (s) Relaxation time= 7.68ms*vd/0.01 xau calibsk for PLDB19 |
