Hi
I am a novice with pombe methodology. So, I need help with deleting a
sequence from the genome. The sequence itself can range between
400-700bp depending upon the strategy used. This sequence in the NCBI
database implies that it is non-coding, since no gene information is
provided. I want to disrupt that sequence with a marker, preferrably
ura4+. All I can think of is to insert ura4+ at the restriction site
and replace ura4+ with a DNA fragment containing the deletion may be
by homologous recombination. But I do not know if it is correct. So,
please send suggestions with a detailed protocol of how I can do that
in an h90 strain.
Thanks,
Sue
