![]() Today's 15% gel (seen above) ran like shit, but yesterday's 15% gel, made at the same time from the same material, was perfect. However I also have no idea what I could be doing wrong that would be causing this issue at this frequency when I have been being absolutely paranoid about consistency since this problem started. They have run hundreds of gels in their career, and as part of trouble shooting this issue this researcher and others when over his methods. I trust the researcher not to be doing anything wrong. It happens to between 1/2 and 2/3rds of the the gels this researcher runs. The smearing seems to be isolated to bands smaller than 25 KDa.Īs mentioned previously, this is not a consistent issue. This may be due to the fact that this researcher is the only person working with proteins below 23 KDa. They, along with three other researchers, run 12.5% and 15% regularly, however they are the only one who experiences this issue even when the gels they're using were poured in the same batch as a gel another researcher used that ran perfectly. The issue consists of vertical smearing of both their sample and the ladder. Right now I have one researcher out of eight who is experiencing a very specific problem with their western blots repeatedly, but not consistently. However this particular problem has me stumped. This makes troubleshooting somewhat difficult, but so far I have managed rather well. Ironically, I have never run a western blot myself, I just make the gels (and do a bunch of other lab minutia that doesn't really matter for this post). On an average day, I make between 8 and 15 gels, with a "slow" day being around 5 and a "busy" day being up to 24. Literally everyone else in the lab uses them, but I am the sole person responsible for making them. I am a lab tech/manager for a lab that runs literally dozens of SDS-PAGE gels/Western Blots a week.
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