Tn-10 translational fusion was analysed by PCR, using the
Ara/LacZ primers that you used on RDM Day 3. A band was
seen with the AraC/LacZ primer set. The AraC/LacZ PCR product
was sequenced in order to confirm that the mutant was indeed
an AraC translational fusion, and to identify precisely
where in ara C the transposon had inserted.
The image on the right is the raw
output from a sequencing gel, using the "dye terminator
method." Eight lanes are shown, representing four reactions
carried out in duplicate (duplicate lanes are right next
to each other, so they may appear as one large lane).
are loaded as follows:
are the same as lanes 5-8, except that they use only
33% of the recommended sequencing kit reagents. "AraC" indicates
that the AraC primer was used to sequence, whereas "LacZ"
samples used the LacZ primer for sequencing.