I assume you have read in the lines and stored them in an array
lines
Then,
set(lines)
gives you a set that contains all unique lines. If every line is unique the length of lines and set(lines) will be the same. Ergo:
if len(lines) == len(set(lines)):
print 'all lines are unique'
else:
print 'not all lines are unique'
solved Finding repeated lines in Python [duplicate]