Question from NMRWiki Q&A forum Running a wet sequence

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[Question from NMRWiki Q&A forum] Running a wet sequence

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NMR processing:

MDD

NMR assignment:

Backbone:

Side-chains:

NOEs:

Structure from NMR restraints:

Ab initio:

Fragment-based:

Rosetta-NMR (Robetta)

Template-based:

GeNMR

I-TASSER

Refinement:

Amber

Structure from chemical shifts:

Fragment-based:

Homology-based:

Torsion angles from chemical shifts:

Preditor

TALOS

Promega- Proline

Secondary structure from chemical shifts:

CSI (via RCI server)

TALOS

MICS caps, β-turns

d2D

PECAN

Flexibility from chemical shifts:

RCI

Interactions from chemical shifts:

HADDOCK

Chemical shifts re-referencing:

NMR model quality:

NOEs, other restraints:

Chemical shifts:

RDCs:

Pseudocontact shifts:

Protein geomtery:

SAVES2 or SAVES4

Quality Control Check

NMR spectrum prediction:

FANDAS

MestReS

V-NMR

Flexibility from structure:

Backbone S2

Methyl S2

B-factor

Molecular dynamics:

Gromacs

Amber

Antechamber

Chemical shifts prediction:

From structure:

Shiftx2

Sparta+

Camshift

CH3shift- Methyl

ArShift- Aromatic

ShiftS

Proshift

PPM

CheShift-2- Cα

From sequence:

Shifty

Camcoil

Poulsen_rc_CS

Disordered proteins:

MAXOCC

Format conversion & validation:

CCPN

From NMR-STAR 3.1

Validate NMR-STAR 3.1

NMR sample preparation:

Protein disorder:

DisMeta

Protein solubility:

Isotope labeling:

Solid-state NMR:

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11-21-2014, 08:58 AM

nmrlearner

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Running a wet sequence

Running a wet sequence

Hi,

I am trying to run a wet sequence but I'm having trouble getting good results on a sample with 90:10 water:methanolD3. O1 is well calibrated as are the 4 selective pulses (I tried both sinc1.100 and Gaus90.1000 shapes) and the gradients. The result is as awful as running a normal proton. In the parameters there are two types of parameters I haven't touched because I don't understand what they do or how they should be calibrated: something called SPOAL (phase alignment of frequency offset) SPOFF (offset frequency) for each of the selective pulses.

I would appreciate any kind of help here.

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