a) How do u tell if your bacteria dna has high GC content? How much of betaine do u usually add to your reaction tube?
If I do use betaine, is it necessary to lower or increase the annealing temperature?
I hope I am thinking too much about the faint band. It is so faint that I stared very long at the image before realizing it Do you usually follow up on an extremely faint band or just rule it out as negative, assuming you do not know whether in first place it is supposed to be present. When I observed the other positive samples I have, they are all very bright clear band. There was one faint like a "pencil line band". Could this be due to high GC content? My product size is about 700+bp I run it on a 1.5% gel.
Usually, would you repeat on such cases?
Any advices very much appreciated!!
Have a great day!
Edited by ocean_sky83, 08 October 2010 - 08:22 AM.