Sauer:HPLC protocol: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
No edit summary |
No edit summary |
||
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. | Under Instrument Set-up, set %B to 0, and T.Flow to 1mL/min. | ||
Line 25: | Line 25: | ||
# 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. | ||
Line 32: | Line 32: | ||
Loading Sample: == | 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: == | 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) |
Revision as of 09:14, 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)