Answer Muscle
The alignment here is a short alignment, we a priori don't know how close related are the species.
- So in this case the defaults parameters should work fine, we do not need to do any modification except for the distance measure that should be change to kmer4_6, as we are working with nucleic acid sequences.
- If you want to run Muscle in a faster way because you are confident about the similarity of your sequences, you can use a limited number of iterations and put a low number in the field:
Maximum number of iterations
, 2 or even 1 should be OK. The anchor optimization with low number of iterations (1-2) has no effect, but you should activate it for more iterations, it speeded the computation and have little or no effect on accuracy. - For huge alignments the first thing you should do is to have a look to the option of
Max. number of hours for align. refinement
, than reduce the number ofMaximum number of iterations
if your sequences are not too divergent, and let the default options for distance measure.