I am trying to sequence a potential KO by submitting samples to Genewiz, and I am getting this repeat in one of my sequences.

Here is my PCR Steps

-98 degrees for 2 minutes

-Touchdown 7 times

-30 Cycles of the following

-98 degree denature for 10 seconds

-60 degree annealing for 20 seconds

-elongation at 72 degrees for 30 seconds

-72 degrees for 5 minutes-

-end cycle

i am not sure if i need to alter my temperature, or if it is my primers, but I am at a loss. does anyone have any ideas?

More Matthew Koslow's questions See All
Similar questions and discussions