Beckman HPLC procedure

 

  1. Check buffer A, buffer B and ACN, make sure not empty.
  2. Check blank solution (H2O) vial not empty. Check makeup buffer vial not empty and is in 85 position.
  3. Open 32 Karat software. Click “Beckman” icon, Click “OK” for instrumental wizard.
  4. Go to Control – Direct Control. Set B as 100% Set flow rate 1ml/min & duration time as 1 min.
  5. 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

  1. Click on blue arrow in the menu bar.
  2. Type down sample ID (this could be the same for a series of runs)
  3. For method, choose “thiolDNA—startup”.
  4. Type down file name (this must be different for each run)
  5. For vial number, type down the number the blank solution vials sits in.
  6. Set injection volume as 5 uL.
  7. Wait until this step is over (Red stop sign on toolbar menu will fade)

 

Sample injection

  1. Make sure makeup buffer is not empty and is on position 85.
  2. Click on blue arrow to start. Choose “thiolDNA-injection” methods. Vial number should be the number your sample is sitting in. And volume is the exact volume of your sample.
  3. Around 20 minutes after, collect the sample from the drain.
  4. 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

  1. Click on blue arrow and choose “thiolDNA-finalclean” method; follow step 9-11.
  2. Pumps and lamps will be turned off automatically.

 

**When changing flow rate or solution percentage manually, always give at least 1 min duration time.