I am running the CometAssay on cryopreserved avian blood samples. I had been obtaining what I thought were good results but then encountered a cloudiness problem the first time I tried to do more than 2 samples (this time 10) at once. I am using two well, pre-made, Trevigen slides and an electrophoresis buffer at pH of 12.1. I have a variety of questions for which I would be grateful for any input!

1) Do you think the sample "S003" images attached look like a good concentration of cells and a good spread of comet tail lengths? I am using OpenComet plugin in ImageJ to automatically score comets.

2) Do you have any ideas about what may have caused the cloudiness in the other images (from samples 008/009) attached? If, you look, it seems like concentric rings formed perhaps as the gel dried (I am sure the gel had dried off the slides completely before staining). I don't really see any tails behind the cells either. The lysis buffer was a bit cloudy during one of my previous runs but not during this latest run. I was wondering if it could have been something with the gel temperature since it took me longer to process 10 samples at once or something wrong with my SYBR Gold stain.

3) The protocol I am following says to run the gel at 21 V and 90 mA, but the voltage source I have requires a higher mA to maintain 21 V. Do you think I should increase the mA to about 250 to allow constant voltage or just keep the settings and accept the voltage staying around only 18?