Sauer:HPLC protocol: Difference between revisions
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== HPLC usage: == | == HPLC usage: == | ||
== | === Buffers: === | ||
Buffers: == | |||
*A: 0.06% TFA in dH2O. | *A: 0.06% TFA in dH2O. | ||
Line 18: | Line 18: | ||
'''Column should be stored in 100% Buffer B.''' | '''Column should be stored in 100% Buffer B.''' | ||
''Flush into Buffer A before run.'' | |||
* Under Instrument Set-up, set %B to 0, and T.Flow to 1mL/min. | |||
'''To run, click Download''' | |||
To run, click Download | |||
# To run a method, open method (start with basic.met, obtain copy and create your own folder for this.) | |||
# To start the method, click on the Blue arrow. | |||
Select a name for your file. Make sure this will be in your own directory. | Select a name for your file. Make sure this will be in your own directory. | ||
'''Loading Sample:''' | |||
Loading Sample: | |||
# Wash the syringe a couple times with Buffer A, shooting the waste into the waste container. | |||
Triggering the Run: | # With the knob turned all the way to the right (clockwise) insert the needle. | ||
# Turn the knob counterclockwise all the way, and then inject your sample. | |||
# Turn the knob back in the clockwise direction. | |||
'''Triggering the Run: ''' | |||
# Press Start on the control box. | |||
# To trigger the run to start, form a circuit between the two exposed wires. | |||
'''Shutdown:''' | |||
Shutdown: | |||
# Go to 100% Buffer B for 10 min. | |||
# Change flowrate to 0. | |||
# Close down Class VP | |||
# Shut-down pumps from top to bottom. | |||
# Clean the Syringe with Buffer A. Leave syringe in HPLC. | |||
(otherwise the next person could end up with your sample as a contaminant) | (otherwise the next person could end up with your sample as a contaminant) |
Latest revision as of 09:18, 23 June 2009
HPLC usage:
Buffers:
- A: 0.06% TFA in dH2O.
- B: 0.054% TFA in 80% Acetonitrile
- When preparing, filter with Chemically Resistant ZapCap.
- Turn on pumps from bottom to top. (there are 4 parts that need to be turned on)
- Uncap waste container
- Turn on computer, open Class VP, open Instrument 1.
- To turn on computer control, click on icon that looks like a stamped letter. (upper left-hand corner.)
Column should be stored in 100% Buffer B.
Flush into Buffer A before run.
- Under Instrument Set-up, set %B to 0, and T.Flow to 1mL/min.
To run, click Download
- To run a method, open method (start with basic.met, obtain copy and create your own folder for this.)
- To start the method, click on the Blue arrow.
Select a name for your file. Make sure this will be in your own directory.
Loading Sample:
- Wash the syringe a couple times with Buffer A, shooting the waste into the waste container.
- With the knob turned all the way to the right (clockwise) insert the needle.
- Turn the knob counterclockwise all the way, and then inject your sample.
- Turn the knob back in the clockwise direction.
Triggering the Run:
- Press Start on the control box.
- To trigger the run to start, form a circuit between the two exposed wires.
Shutdown:
- Go to 100% Buffer B for 10 min.
- Change flowrate to 0.
- Close down Class VP
- Shut-down pumps from top to bottom.
- Clean the Syringe with Buffer A. Leave syringe in HPLC.
(otherwise the next person could end up with your sample as a contaminant)