DNA Ladder Issues
Posted 06 November 2009 - 12:51 PM
I am having an issue with my 100bp DNA ladder on my Agarose Gel (1.5%). The heavier bands of the ladder (500 and up) do not separate and leave a blurry mess. I moved the ladder to the middle of the gel and reduced the concentration by half. The ladder got slightly clearer but still not readable. I need to be able to differentiate between 750bp and 1000 bp for my PCR product. I cast and run the gel in 0.5X TBE and I don't reuse the buffer more than 2 or 3 times. I attached a pic of a gel with the ladder problem.
I am very new at this so I would appreciate any suggestions you might have.
Posted 06 November 2009 - 11:10 PM
from your pictire its clear that the gel is just half of it.. try runnign the entire gel.. i bet my money on it u l get the required resolution!!!
higher mol wt molecules take longer time to resolve owing to the steric hinderences too!!
Posted 09 November 2009 - 12:03 AM
Posted 09 November 2009 - 04:23 AM
I run 140V for 30 minutes.
Hope this helps you.
Edited by adrian kohsf, 09 November 2009 - 04:25 AM.
..."best of our knowledge, as far as we know this had never been reported before, though I can't possible read all the published journals on earth, but by perform thorough search in google, the keywords did not match any documents"...
"what doesn't kill you, makes you stronger"---Goddess Casandra reminds me to be strong
"It's all just DNA. Do it."---phage434
Posted 09 November 2009 - 07:36 AM