Common problems to avoid when running PAGE gel electrophoresis:
* Forgetting to remove tape sealing precast gels
* green strip on bottom of BIORAD gels, white strip on the side of Novex gels
* Not ensuring the gasket is tight
* for the BIORAD tetra cell boxes, make sure the edge of the inner plate (or top of the ledge of the buffer dam) is snug up against the bottom (not the side) of the green ledge on the rubber seal)
* pour buffer into the inner chamber first, wait to see if it leaks - if it does, redo; if it doesn’t, pour buffer into outer chamber
* Having too little buffer
* make sure that the buffer level in the outer chamber at least covers the level of the wire & that the inner chamber is full
* Overloading lanes
* typically don’t want to run more than 2 ug of purified protein or 20 ug total protein in a lysate similar on a typical SDS-PAGE mini gel
* Not having good contact between the electrodes and the lid
* if running 4 gels in a tetra cell, you may need to bend the metal plates on the bottom of the lid so that they make contact with the screws
* Swapping the electrodes and/or putting your samples at the wrong end
* harder to do with PAGE gels than agarose ones, but it is possible! so make sure you match red to red and black to black
more gel electrophoresis videos: https://www.youtube.com/playlist?list=PLUWsCDtjESrEzmik3IyuOAYrPkisVbewS
more practical lab tips: https://bit.ly/lab_tricks_page & https://www.youtube.com/playlist?list=PLUWsCDtjESrFEAWZCRKJL7sMc6a_KgfLU
more about all sorts of things: #365DaysOfScience All (with topics listed) 👉 http://bit.ly/2OllAB0 or search blog: http://thebumblingbiochemist.com