Beckman HPLC procedure
- Check
buffer A, buffer B and ACN, make sure not empty.
- Check
blank solution (H2O) vial not empty. Check makeup buffer vial
not empty and is in 85 position.
- Open
32 Karat software. Click ÒBeckmanÓ icon, Click ÒOKÓ for instrumental wizard.
- Go to
Control Ð Direct Control. Set B as 100% Set flow rate 1ml/min &
duration time as 1 min.
- Turn
on Lamp by clicking the lamp icon at the right side, and wash column by
clicking the syringe icon in the middle. This only needs to be done once
at startup.
Start up program
- Click
on blue arrow in the menu bar.
- Type
down sample ID (this could be the same for a series of runs)
- For
method, choose ÒthiolDNAÑstartupÓ.
- Type
down file name (this must be different for each run)
- For
vial number, type down the number the blank solution vials sits in.
- Set
injection volume as 5 uL.
- Wait
until this step is over (Red stop sign on toolbar menu will fade)
Sample injection
- Make
sure makeup buffer is not empty and is on position 85.
- Click
on blue arrow to start. Choose ÒthiolDNA-injectionÓ methods. Vial number
should be the number your sample is sitting in. And volume is the exact
volume of your sample.
- Around
20 minutes after, collect the sample from the drain.
- If you
have more samples to inject, just wait until the program finishes, then go
to step 17; otherwise stop the program by clicking the stop sign and set
B% to 10% at direct control window.
Final cleaning
- Click
on blue arrow and choose ÒthiolDNA-finalcleanÓ method; follow step 9-11.
- Pumps
and lamps will be turned off automatically.
**When changing flow rate or
solution percentage manually, always give at least 1 min duration time.