User:Stuart McKellar/Notebook/Bird Sex Testing/2012/11/14
|Project name|| Main project page|
Previous entry Next entry
Checking DNA in samples
The aim is to see if there is still DNA in my samples, pre-PCR. The ladder was added as a positive control to check if the gel was still able to stain the DNA.
ran gel @ 300V, 60mA. No melt. Both DNA ladders were visible. The lanes contained 5ul sample and 1ul loading buffer. No visible DNA in the lanes besides the ladder. I modified the TI back to a standalone form. There should be a ton of DNA in the PCR samples too. There may not be enough contrast to see the PCR samples so I will try it in a dark room, wash the gel, and also will try a post stain maybe. This is hard to do as i don't have any GelGreen left.
Checked the gel in the dark and it showed faint bands in almost all the lanes. One band had a very faint single band. Others showed many bands, suggesting non-specific binding of primers. AMG suggested raising the temperature of the annealing step as this will reduce non-specific binding. She also advised that there could be contamination. The faint bands could also be due to non-specific binding. Steps to take from here: