Collecting a 15N Edited HSQC - Revision history

Mjeppesen at 20:30, 21 July 2022

2022-07-21T20:30:40Z

← Older revision		Revision as of 20:30, 21 July 2022
Line 23:		Line 23:
	# Rule of thumb - If you can collect an amazing NHSQC in 10 min you are ready for 3D work.		# Rule of thumb - If you can collect an amazing NHSQC in 10 min you are ready for 3D work.

	~~[[category:Protocols]]~~
	[[category:Data_Collection]]		[[category:Data_Collection]]
	[[category:NMR_Usage]]		[[category:NMR_Usage]]

Mjeppesen: LearningSorting

2021-09-11T04:53:54Z

LearningSorting

← Older revision		Revision as of 04:53, 11 September 2021
Line 23:		Line 23:
	# Rule of thumb - If you can collect an amazing NHSQC in 10 min you are ready for 3D work.		# Rule of thumb - If you can collect an amazing NHSQC in 10 min you are ready for 3D work.

	[[category:~~Protocols\|~~Protocols]]		[[category:Protocols]]
	[[category:Data_Collection]]		[[category:Data_Collection]]
			[[category:NMR_Usage]]

Mjeppesen: Added to Category

2021-09-11T03:18:41Z

Added to Category

← Older revision		Revision as of 03:18, 11 September 2021
Line 22:		Line 22:
	# Edit phasing manually with ‘ph’.		# Edit phasing manually with ‘ph’.
	# Rule of thumb - If you can collect an amazing NHSQC in 10 min you are ready for 3D work.		# Rule of thumb - If you can collect an amazing NHSQC in 10 min you are ready for 3D work.

			[[category:Protocols\|Protocols]]
			[[category:Data_Collection]]

Jcatazaro: Created page with "Collecting a Superb 15N edited HSQC (NHSQC) # Concentrate your protein as much as possible. It should be at least 50uM for 2D work and around 1mM for 3D work. # Spike in 10% ..."

2017-03-14T21:16:01Z

Created page with "Collecting a Superb 15N edited HSQC (NHSQC) # Concentrate your protein as much as possible. It should be at least 50uM for 2D work and around 1mM for 3D work. # Spike in 10% ..."

New page

Collecting a Superb 15N edited HSQC (NHSQC)

# Concentrate your protein as much as possible. It should be at least 50uM for 2D work and around 1mM for 3D work.
# Spike in 10% D2O.
# Place in a Shigemi tube (if low volume) or in any other NMR tube. Cap and seal.
# Insert your sample into the spectrometer. Create a new dataset.
# Open experimental parameter set ‘NHSQC’. If you can’t find it, open hsqcetf3gpsi. Also, double check that this is the pulse program.
# Edit the number of points in the direct and indirect dimension. Should be 2k and >= 128.
# The number of dummy scan should be at least 16.
# The number of scans should be adjusted depending on the concentration. For 1mM protein, 2 scans is ok. For 500uM protein, 8 scans will be necessary.
# Edit your sweep width in the indirect dimension. A larger sweep width will prevent folding of the spectrum but will decrease digital resolution.
# Edit your 15N center position. It should be between 115-120 ppm.
# Edit the ZGOPTNS. Add –DLABEL_CN if the protein is double labeled. This will tell the pulse program to decouple 13C.
# Check experiment time with ‘expt’.
# Lock ‘lock h2o+d2o’.
# Tune ‘atma’.
# Shim ‘topshim’ or ‘topshim 3d’.
# Find p1 ‘pulsecal’.
# Calculate receiver gain ‘rga’.
# Zero go ‘zg’.
# Process the specrum with ‘xfb’.
# Edit phasing manually with ‘ph’.
# Rule of thumb - If you can collect an amazing NHSQC in 10 min you are ready for 3D work.