
DNA sequencing
ladder
Parallel
electrophoresis of the four ddNTP
termination reactions produces a "ladder" in which each "rung"
is
a fragment one nucleotide longer than the one below it, either
in
the
same or a different lane. By labeling the lanes of the gel
with
the
letter
of the base complementary
to
the ddNTP
reaction loaded, the sequence of
the original DNA strand can be read directly in the 5'
3'
direction, by moving from
bottom
to top of the autoradogram.
HOMEWORK:
In a typical experiment, both strands of a dsDNA molecule
are sequenced and compared as cross-checks. As both strands are
read 5'
3' direction, direct alignment of the
two sequences is difficult. Can you suggest a simple means to
read the second autoradiogram with the base complementary
sequence, so that the reverse strand
sequence reads the same as the forward strand
sequence, in the same 5'
3' direction?