Last time I had finished up all of my UV/Vis and Fluroescence spectral work in addition to developing and creating my method for capillary electrophoresis separation. I finally got the chance to test out my method on the Bio-Rad CE here in the lab! In order to do that, I had to make my capillary by measuring it to size, cutting gently, feeding it through the CE cartridge, and trimming the ends before inserting into the instrument. Next, I filled my vials to place in the CE (inlet/outlet buffer, 1.0 M NaOH rinse, water rinse, buffer rinse, and waste). I was ready to begin – vials ready? (check), sample ready? (check), capillary inserted? (check), method developed? (check), laser on? (check)– READY, SET, GO!
For the past three days, I altered my method by changing the injection pressure, voltage, and concentration of the sample; however, my separations were just not providing consistent, clear results. No reproducibility! Kathryn suggested running an electric diagnostic test after the current reached maximum and aborted the separation a few times. The electric diagnostic proved we could not go above 18kV – not as one would expect. It was time to consult Dr. Colyer and go to plan B!
Just as Thomas Edison tried 1,000 times to invent the incandescent light bulb, he kept trying. Thomas Edison stated, “Our greatest weakness lies in giving up. The most certain way to succeed is always to try just one more time.” This quote defines research – one must not give up with a test is not successful, but try one more time and the answer may lie ahead.
Check back for what Plan B will shed light on! (Haha, I just had to.)