Initially when we first started using Gel (2003) we were taught to hold the pipetor at a 30-45 degree angle to dispense the 0.8% cells and plasma into each well of the Anti-IgG Cards. Since then we have had several questions arise about Gel procedures. The technical support staff at Ortho told me on two separate occasions that there is no actual documented procedure as to how to hold the pipetor when dispensing. I was also told that the you should try to keep the bubble intact, but as long as the cell suspension has not descended into the gel before centrifugation it will be ok, but it may decrease if the bubble gets dislodged and separates the sample so disproportionate amounts of cells and plasma get trapped on either side.(I've found that it does sometimes affect the strength of reactivity that you see when weaker antibodies are present.) but it is definitely not optimal, and I've trained my staff that if the top of the gel is not touching the cells that they just added, then it is fine. If it is touching the top of the gel then they are to repeat it in another well of the cards and not to use the reactions in those wells that weren't pipetted correctly.
