DNA molecular weight keeps changes
Posted 24 September 2010 - 08:23 AM
What could be wrong? Not only are the MW changing, the higher MW bands are poorly resolved/smearing. I'm using Fermentas 1kb GeneRuler. Instead of using 1uL (500ng), I premixed the loading dye and ladder such that I use only 1/5 the recommended amount (100ng). Could premixing be the problem (eg dye interference)? I've done this at my last lab with NEB 1kb ladder with no problem. I tried a new tube of ladder and it's still the same. But then, I used SB and ethidium bromide there instead of TAE and GelStar. Stains are added to the gel. I've tried preparing new buffers from commercial 10x TAE and making new gels. I'm running at 100V (~5V/cm). Is that too high?
Please help. Thanks!
Posted 24 September 2010 - 09:54 PM
Posted 25 September 2010 - 01:28 AM
Edited by donny, 25 September 2010 - 01:29 AM.
Posted 25 September 2010 - 02:53 PM