I am trying to develop a method to synthesize 100-200 bases of ssDNA product from PCR. I have a template lambda DNA. presently using assymetric pcr i am getting more than one strand in denaturng PAGE. To develop my method do i need to use a shorter template as i can reduce the mispriming and unwanted products.
I am using denaturing PAGE gels of 15%, but normal dna ladders 100 bp dont run as i have urea in gel. Can i use a rna ladder.
if have to make this method work with a normal pcr protocol will it be possible. what do i need to do to make my gels work.
I m relatively new to this filed so I am asking this question.
sorry for the long question and thanks in advance for any suggestion.