Try removing the following two lines from your board() function and placing them between num_players and board() at the bottom of your script like so:
Num_Humans()
root = Tkinter.Tk()
root.attributes('-topmost', True)
Board()
I hope that's what you were looking for
EDIT:
As far as tips here's my first:
The 50 lines of code used to generate the T1 through T50 labels can be simplified to the following two lines:
for i in xrange( 1, 51 ):
exec 'T%s = Tkinter.Label(root, text ="%s", relief=RIDGE,padx=3,pady=10 )' % (i,i)
This can be used for the other long sections of code, but this is the most apparent need for it...