LOCAL

Lab Protocols

 Proper Loading of Sample in AKTA

 

 First, a definition of terms:

 "Loading" is the process of loading the sample into the loop--the
 operator does this part manually in general.

 "Injecting" is the process of the sample's being pumped from the loop
 and into the column. The AKTA pumps take care of this part.

 Confusion of these two terms is likely a major reason that
 proper sample loading technique is frequently misunderstood.

 To properly load sample from a syringe to the AKTA:

   1. Clear all air bubbles from the sample in the syringe
      (to protect sizing columns).
   2. GENTLY insert the syringe into the syringe valve until
      it stops. As Dan might say, you only need 0.001% finger
      strength to do this. DO NOT press the syringe in as if you are
      trying to puncture something. The seal is formed by the
      blunt abutting of two surfaces, NOT by the pressure
      of a punctured membrane surrounding the needle.
      The surfaces only need to make contact to form a
      seal. The needle does not puncture anything.
   3. Leave the syringe and needle in place until the
      sample is injected and the valve position moves
      back to "load" or else you risk losing your
      sample to gravity.
   4. NEVER use a "syringe needle" with a pointed tip to
      load the AKTA. The question you want to ask yourself
      when loading the AKTA is: "Would I want what I am about to
      do done to my inner ear?"